Experimental Design Schematic

Fig.1 Experimental design showing treatments and husbandry events during the course of the study. Symbols indicate when an experimental event occurred at each time point. (1) 260 fish were assigned and acclimated to one of three water temperature groups (e.g., 28°C, 32°C, or 35°C) and reared from 0 to 164 days post fertilization (dpf). (2a) At 164 dpf (or 0 days post exposure; DPE), fecal collections were collected from a random selection of five fish per tank (n = 60). Additionally, histological and wet mount assessments were conducted on selected fish to assess presence of infection and infection burden. (1b) Afterwards, a cohort of fish from each water temperature group were exposed to Pseudocapillaria tomentosa. (3-6) Subsequent fecal samples were collected and histopathological assessments were conducted at 14 dpe (n = 54), (3) 21 dpe (n = 48), (4) 28 dpe (n = 47), and (5) 42 dpe (n = 51).

Results

2) Water temperature shapes gut microbiome structure

Fig. 2 Effects of water temperature on gut microbiomes of parasite unexposed zebrafish. (A) Simpson’s Index of diversity shows that gut microbiome diversity significantly differs between fish reared at 28°C and 35°C water temperatures. (B) Capscale ordination based on the Bray-Curtis dissimilarity of gut microbiome composition. The analysis shows that gut microbiome composition significantly differs between fish reared at different water temperatures. (C) Simpson’s Index of diversity shows microbial gut diversity increases with time from 0 days post exposure (dpe) to 42 dpe, irrespective of water temperature. (D) Capscale ordination of gut microbiome composition based on the Bray-Curtis dissimilarity constrained on the main effects of water temperature and time, and their interaction. The analysis shows that shows that gut microbiome composition differs between fish across time depending on water temperature. Ribbons and ellipses indicate 95% confidence interval. Only statistically significant relationships are shown. A “*” indicates statistical significance below the “0.05” level. Black arrows indicate direction of greatest change in the indicated covariates.

2A)

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GLM
GLM Results
glm(Alpha.Score ~ Temperature); Unexposed fish
term estimate std.error statistic p.value p.adj.sig
Shannon
(Intercept) −0.754 0.091 −8.309 <0.001 ****
Temperature32 0.051 0.127 0.400 ≥0.25 ns
Temperature35 0.054 0.129 0.421 ≥0.25 ns
Simpson
(Intercept) −0.101 0.092 −1.103 ≥0.25 ns
Temperature32 0.132 0.128 1.030 ≥0.25 ns
Temperature35 0.340 0.131 2.591 0.010 *
Richness
(Intercept) −0.231 0.075 −3.079 0.002 **
Temperature32 0.047 0.105 0.446 ≥0.25 ns
Temperature35 −0.473 0.110 −4.299 <0.001 ****
ANOVA
ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature), type = 2); Unexposed fish
term statistic df p.value sig
Shannon
Temperature 0.225 2.000 ≥0.25 ns
Simpson
Temperature 6.830 2.000 0.033 *
Richness
Temperature 27.512 2.000 <0.001 ****
Tukey
Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature); Unexposed fish
term .y. group1 group2 estimate std.error statistic adj.p.value Variable
Shannon
Temperature Alpha.Score 32 28 0.051 0.127 0.400 ≥0.25 Temperature
Temperature Alpha.Score 35 28 0.054 0.129 0.421 ≥0.25 Temperature
Temperature Alpha.Score 35 32 0.004 0.127 0.029 ≥0.25 Temperature
Simpson
Temperature Alpha.Score 32 28 0.132 0.128 1.030 ≥0.25 Temperature
Temperature Alpha.Score 35 28 0.340 0.131 2.591 0.026 Temperature
Temperature Alpha.Score 35 32 0.207 0.130 1.596 0.247 Temperature
Richness
Temperature Alpha.Score 32 28 0.047 0.105 0.446 ≥0.25 Temperature
Temperature Alpha.Score 35 28 −0.473 0.110 −4.299 <0.001 Temperature
Temperature Alpha.Score 35 32 −0.520 0.109 −4.768 <0.001 Temperature

2B)

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Capscale
ADONIS2
adonis2(Beta Distance ~ Temperature); Unexposed fish
term df SumOfSqs R2 statistic p.value sig
bray
Temperature 2.000 2.294 0.133 12.900 0.001 ***
Residual 168.000 14.936 0.867 NA NA NA
Total 170.000 17.229 1.000 NA NA NA
canberra
Temperature 2.000 3.862 0.099 9.178 0.001 ***
Residual 168.000 35.344 0.901 NA NA NA
Total 170.000 39.206 1.000 NA NA NA
gunifrac
Temperature 2.000 1.422 0.102 9.566 0.001 ***
Residual 168.000 12.485 0.898 NA NA NA
Total 170.000 13.907 1.000 NA NA NA
Dispersion (ANOVA)
ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature); Unexposed fish
term df sumsq meansq statistic p.value sig
bray
Temperature 2.000 0.008 0.004 0.386 ≥0.25 ns
Residual 168.000 1.771 0.011 NA NA NA
canberra
Temperature 2.000 0.008 0.004 0.873 ≥0.25 ns
Residual 168.000 0.759 0.005 NA NA NA
gunifrac
Temperature 2.000 0.005 0.003 0.369 ≥0.25 ns
Residual 168.000 1.143 0.007 NA NA NA
Dispersion (Tukey)
Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature); Unexposed fish
.y. term group1 group2 estimate conf.low conf.high adj.p.value sig
bray
Distance Temperature 32 28 −0.011 −0.056 0.034 ≥0.25 ns
Distance Temperature 35 28 0.005 −0.041 0.051 ≥0.25 ns
Distance Temperature 35 32 0.016 −0.029 0.062 ≥0.25 ns
canberra
Distance Temperature 32 28 −0.001 −0.031 0.028 ≥0.25 ns
Distance Temperature 35 28 0.014 −0.016 0.044 ≥0.25 ns
Distance Temperature 35 32 0.015 −0.015 0.045 ≥0.25 ns
gunifrac
Distance Temperature 32 28 −0.013 −0.049 0.023 ≥0.25 ns
Distance Temperature 35 28 −0.004 −0.040 0.033 ≥0.25 ns
Distance Temperature 35 32 0.009 −0.027 0.046 ≥0.25 ns

2C)

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GLM
GLM Results
glm(Alpha.Score ~ Temperature*DPE); Unexposed fish
term estimate std.error statistic p.value p.adj.sig
Shannon
(Intercept) −1.074 0.138 −7.754 <0.001 ****
Temperature32 0.075 0.193 0.386 ≥0.25 ns
Temperature35 0.154 0.193 0.797 ≥0.25 ns
DPE 0.018 0.006 3.086 0.002 **
Temperature32:DPE −0.001 0.008 −0.170 ≥0.25 ns
Temperature35:DPE −0.005 0.008 −0.585 ≥0.25 ns
Simpson
(Intercept) −0.433 0.134 −3.245 0.001 **
Temperature32 0.157 0.186 0.841 ≥0.25 ns
Temperature35 0.408 0.187 2.177 0.031 *
DPE 0.019 0.006 3.293 0.001 **
Temperature32:DPE −0.001 0.008 −0.173 ≥0.25 ns
Temperature35:DPE −0.003 0.008 −0.343 ≥0.25 ns
Richness
(Intercept) −0.651 0.108 −6.017 <0.001 ****
Temperature32 0.189 0.150 1.257 0.211 ns
Temperature35 −0.059 0.154 −0.383 ≥0.25 ns
DPE 0.024 0.005 5.137 <0.001 ****
Temperature32:DPE −0.008 0.006 −1.251 0.213 ns
Temperature35:DPE −0.024 0.007 −3.476 <0.001 ***
ANOVA
ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature*Time), type = 2); Unexposed fish
term statistic df p.value sig
Shannon
Temperature 0.322 2.000 ≥0.25 ns
DPE 22.895 1.000 <0.001 ****
Temperature:DPE 0.363 2.000 ≥0.25 ns
Simpson
Temperature 8.551 2.000 0.014 *
DPE 28.161 1.000 <0.001 ****
Temperature:DPE 0.118 2.000 ≥0.25 ns
Richness
Temperature 31.759 2.000 <0.001 ****
DPE 27.201 1.000 <0.001 ****
Temperature:DPE 12.460 2.000 0.002 **
Tukey
Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature*Time); Unexposed fish
Temperature .y. term group1 group2 estimate std.error statistic adj.p.value Variable Group
Shannon
28 Alpha.Score DPE 14 0 1.065 0.294 3.625 0.003 DPE 28
28 Alpha.Score DPE 21 0 0.722 0.281 2.566 0.076 DPE 28
28 Alpha.Score DPE 28 0 1.187 0.274 4.335 <0.001 DPE 28
28 Alpha.Score DPE 42 0 0.548 0.296 1.853 ≥0.25 DPE 28
28 Alpha.Score DPE 21 14 −0.343 0.317 −1.083 ≥0.25 DPE 28
28 Alpha.Score DPE 28 14 0.122 0.310 0.392 ≥0.25 DPE 28
28 Alpha.Score DPE 42 14 −0.517 0.330 −1.567 ≥0.25 DPE 28
28 Alpha.Score DPE 28 21 0.465 0.299 1.558 ≥0.25 DPE 28
28 Alpha.Score DPE 42 21 −0.174 0.319 −0.545 ≥0.25 DPE 28
28 Alpha.Score DPE 42 28 −0.639 0.312 −2.045 0.243 DPE 28
32 Alpha.Score DPE 14 0 0.006 0.250 0.022 ≥0.25 DPE 32
32 Alpha.Score DPE 21 0 0.112 0.247 0.452 ≥0.25 DPE 32
32 Alpha.Score DPE 28 0 0.420 0.248 1.695 ≥0.25 DPE 32
32 Alpha.Score DPE 42 0 0.694 0.236 2.943 0.027 DPE 32
32 Alpha.Score DPE 21 14 0.106 0.286 0.371 ≥0.25 DPE 32
32 Alpha.Score DPE 28 14 0.414 0.287 1.445 ≥0.25 DPE 32
32 Alpha.Score DPE 42 14 0.688 0.276 2.490 0.092 DPE 32
32 Alpha.Score DPE 28 21 0.308 0.283 1.087 ≥0.25 DPE 32
32 Alpha.Score DPE 42 21 0.582 0.273 2.131 0.205 DPE 32
32 Alpha.Score DPE 42 28 0.274 0.274 1.001 ≥0.25 DPE 32
35 Alpha.Score DPE 14 0 −0.006 0.210 −0.027 ≥0.25 DPE 35
35 Alpha.Score DPE 21 0 −0.249 0.218 −1.145 ≥0.25 DPE 35
35 Alpha.Score DPE 28 0 0.232 0.204 1.138 ≥0.25 DPE 35
35 Alpha.Score DPE 42 0 0.661 0.207 3.194 0.012 DPE 35
35 Alpha.Score DPE 21 14 −0.243 0.253 −0.962 ≥0.25 DPE 35
35 Alpha.Score DPE 28 14 0.238 0.242 0.985 ≥0.25 DPE 35
35 Alpha.Score DPE 42 14 0.667 0.244 2.733 0.049 DPE 35
35 Alpha.Score DPE 28 21 0.481 0.248 1.938 ≥0.25 DPE 35
35 Alpha.Score DPE 42 21 0.910 0.251 3.628 0.003 DPE 35
35 Alpha.Score DPE 42 28 0.429 0.239 1.792 ≥0.25 DPE 35
Simpson
28 Alpha.Score DPE 14 0 1.256 0.273 4.611 <0.001 DPE 28
28 Alpha.Score DPE 21 0 0.761 0.247 3.076 0.018 DPE 28
28 Alpha.Score DPE 28 0 1.334 0.254 5.241 <0.001 DPE 28
28 Alpha.Score DPE 42 0 0.514 0.257 2.002 ≥0.25 DPE 28
28 Alpha.Score DPE 21 14 −0.496 0.302 −1.644 ≥0.25 DPE 28
28 Alpha.Score DPE 28 14 0.077 0.308 0.251 ≥0.25 DPE 28
28 Alpha.Score DPE 42 14 −0.742 0.310 −2.398 0.114 DPE 28
28 Alpha.Score DPE 28 21 0.573 0.285 2.008 ≥0.25 DPE 28
28 Alpha.Score DPE 42 21 −0.246 0.288 −0.857 ≥0.25 DPE 28
28 Alpha.Score DPE 42 28 −0.819 0.294 −2.789 0.042 DPE 28
32 Alpha.Score DPE 14 0 0.065 0.249 0.261 ≥0.25 DPE 32
32 Alpha.Score DPE 21 0 0.142 0.249 0.569 ≥0.25 DPE 32
32 Alpha.Score DPE 28 0 0.592 0.261 2.272 0.152 DPE 32
32 Alpha.Score DPE 42 0 0.724 0.254 2.850 0.035 DPE 32
32 Alpha.Score DPE 21 14 0.077 0.287 0.267 ≥0.25 DPE 32
32 Alpha.Score DPE 28 14 0.527 0.297 1.773 ≥0.25 DPE 32
32 Alpha.Score DPE 42 14 0.659 0.292 2.260 0.156 DPE 32
32 Alpha.Score DPE 28 21 0.451 0.297 1.517 ≥0.25 DPE 32
32 Alpha.Score DPE 42 21 0.582 0.291 1.999 ≥0.25 DPE 32
32 Alpha.Score DPE 42 28 0.132 0.302 0.436 ≥0.25 DPE 32
35 Alpha.Score DPE 14 0 0.149 0.219 0.683 ≥0.25 DPE 35
35 Alpha.Score DPE 21 0 −0.167 0.218 −0.766 ≥0.25 DPE 35
35 Alpha.Score DPE 28 0 0.413 0.222 1.858 ≥0.25 DPE 35
35 Alpha.Score DPE 42 0 0.836 0.244 3.431 0.005 DPE 35
35 Alpha.Score DPE 21 14 −0.316 0.257 −1.232 ≥0.25 DPE 35
35 Alpha.Score DPE 28 14 0.263 0.260 1.013 ≥0.25 DPE 35
35 Alpha.Score DPE 42 14 0.687 0.279 2.465 0.097 DPE 35
35 Alpha.Score DPE 28 21 0.579 0.260 2.233 0.165 DPE 35
35 Alpha.Score DPE 42 21 1.003 0.278 3.604 0.003 DPE 35
35 Alpha.Score DPE 42 28 0.424 0.282 1.506 ≥0.25 DPE 35
Richness
28 Alpha.Score DPE 14 0 0.432 0.236 1.836 ≥0.25 DPE 28
28 Alpha.Score DPE 21 0 0.550 0.218 2.526 0.084 DPE 28
28 Alpha.Score DPE 28 0 0.767 0.218 3.525 0.004 DPE 28
28 Alpha.Score DPE 42 0 0.970 0.227 4.278 <0.001 DPE 28
28 Alpha.Score DPE 21 14 0.118 0.263 0.449 ≥0.25 DPE 28
28 Alpha.Score DPE 28 14 0.335 0.262 1.275 ≥0.25 DPE 28
28 Alpha.Score DPE 42 14 0.537 0.270 1.990 ≥0.25 DPE 28
28 Alpha.Score DPE 28 21 0.217 0.247 0.880 ≥0.25 DPE 28
28 Alpha.Score DPE 42 21 0.419 0.255 1.647 ≥0.25 DPE 28
28 Alpha.Score DPE 42 28 0.203 0.254 0.796 ≥0.25 DPE 28
32 Alpha.Score DPE 14 0 −0.170 0.177 −0.965 ≥0.25 DPE 32
32 Alpha.Score DPE 21 0 −0.026 0.175 −0.147 ≥0.25 DPE 32
32 Alpha.Score DPE 28 0 0.187 0.180 1.041 ≥0.25 DPE 32
32 Alpha.Score DPE 42 0 0.769 0.176 4.359 <0.001 DPE 32
32 Alpha.Score DPE 21 14 0.145 0.204 0.711 ≥0.25 DPE 32
32 Alpha.Score DPE 28 14 0.358 0.208 1.721 ≥0.25 DPE 32
32 Alpha.Score DPE 42 14 0.939 0.205 4.585 <0.001 DPE 32
32 Alpha.Score DPE 28 21 0.213 0.206 1.031 ≥0.25 DPE 32
32 Alpha.Score DPE 42 21 0.794 0.203 3.906 <0.001 DPE 32
32 Alpha.Score DPE 42 28 0.581 0.208 2.801 0.040 DPE 32
35 Alpha.Score DPE 14 0 −0.560 0.214 −2.612 0.067 DPE 35
35 Alpha.Score DPE 21 0 −0.551 0.214 −2.578 0.073 DPE 35
35 Alpha.Score DPE 28 0 0.110 0.196 0.561 ≥0.25 DPE 35
35 Alpha.Score DPE 42 0 0.007 0.206 0.036 ≥0.25 DPE 35
35 Alpha.Score DPE 21 14 0.008 0.259 0.032 ≥0.25 DPE 35
35 Alpha.Score DPE 28 14 0.670 0.245 2.733 0.048 DPE 35
35 Alpha.Score DPE 42 14 0.567 0.253 2.240 0.162 DPE 35
35 Alpha.Score DPE 28 21 0.661 0.245 2.702 0.053 DPE 35
35 Alpha.Score DPE 42 21 0.559 0.253 2.210 0.173 DPE 35
35 Alpha.Score DPE 42 28 −0.103 0.238 −0.431 ≥0.25 DPE 35

2D)

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Capscale
ADONIS2
adonis2(Beta Distance ~ Temperature*DPE); Unexposed fish
term df SumOfSqs R2 statistic p.value sig
bray
Temperature 2.000 2.294 0.133 16.443 0.001 ***
DPE 1.000 2.531 0.147 36.284 0.001 ***
Temperature:DPE 2.000 0.896 0.052 6.425 0.001 ***
Residual 165.000 11.509 0.668 NA NA NA
Total 170.000 17.229 1.000 NA NA NA
canberra
Temperature 2.000 3.862 0.099 10.667 0.001 ***
DPE 1.000 3.655 0.093 20.192 0.001 ***
Temperature:DPE 2.000 1.820 0.046 5.026 0.001 ***
Residual 165.000 29.869 0.762 NA NA NA
Total 170.000 39.206 1.000 NA NA NA
gunifrac
Temperature 2.000 1.422 0.102 11.984 0.001 ***
DPE 1.000 2.123 0.153 35.785 0.001 ***
Temperature:DPE 2.000 0.574 0.041 4.840 0.001 ***
Residual 165.000 9.788 0.704 NA NA NA
Total 170.000 13.907 1.000 NA NA NA
Dispersion (ANOVA)
ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature); Unexposed fish
term df sumsq meansq statistic p.value sig
bray
Temp.DPE 14.000 0.290 0.021 2.348 0.006 **
Residual 156.000 1.376 0.009 NA NA NA
canberra
Temp.DPE 14.000 0.250 0.018 3.344 <0.001 ***
Residual 156.000 0.831 0.005 NA NA NA
gunifrac
Temp.DPE 14.000 0.249 0.018 2.986 <0.001 ***
Residual 156.000 0.929 0.006 NA NA NA
Dispersion (Tukey)
Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature*DPE); Unexposed fish
.y. term group1 group2 estimate conf.low conf.high adj.p.value sig
bray
Distance Temp.DPE 28°C__14DPE 28°C__0DPE 0.024 −0.112 0.159 ≥0.25 ns
Distance Temp.DPE 28°C__21DPE 28°C__0DPE 0.052 −0.074 0.177 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__0DPE 0.007 −0.118 0.133 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__0DPE −0.017 −0.147 0.113 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__0DPE 0.097 −0.005 0.199 0.085 ns
Distance Temp.DPE 32°C__14DPE 28°C__0DPE 0.041 −0.085 0.166 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__0DPE −0.018 −0.143 0.108 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__0DPE 0.028 −0.102 0.158 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__0DPE 0.017 −0.108 0.142 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__0DPE 0.044 −0.058 0.146 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__0DPE 0.004 −0.126 0.134 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__0DPE −0.004 −0.134 0.126 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__0DPE 0.033 −0.097 0.162 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__0DPE 0.146 0.011 0.282 0.021 *
Distance Temp.DPE 28°C__21DPE 28°C__14DPE 0.028 −0.126 0.181 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__14DPE −0.016 −0.170 0.137 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__14DPE −0.040 −0.198 0.117 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__14DPE 0.073 −0.062 0.209 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__14DPE 0.017 −0.136 0.171 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__14DPE −0.041 −0.195 0.112 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__14DPE 0.005 −0.153 0.162 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__14DPE −0.006 −0.160 0.147 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__14DPE 0.020 −0.115 0.156 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__14DPE −0.019 −0.176 0.138 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__14DPE −0.028 −0.185 0.129 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__14DPE 0.009 −0.148 0.166 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__14DPE 0.123 −0.039 0.284 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__21DPE −0.044 −0.189 0.101 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__21DPE −0.068 −0.217 0.081 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__21DPE 0.045 −0.080 0.171 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__21DPE −0.011 −0.156 0.134 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__21DPE −0.069 −0.214 0.076 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__21DPE −0.023 −0.172 0.125 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__21DPE −0.034 −0.179 0.110 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__21DPE −0.008 −0.133 0.118 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__21DPE −0.047 −0.196 0.102 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__21DPE −0.056 −0.205 0.093 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__21DPE −0.019 −0.168 0.130 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__21DPE 0.095 −0.059 0.248 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__28DPE −0.024 −0.173 0.125 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__28DPE 0.089 −0.036 0.215 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__28DPE 0.033 −0.111 0.178 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__28DPE −0.025 −0.170 0.120 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__28DPE 0.021 −0.128 0.169 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__28DPE 0.010 −0.135 0.154 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__28DPE 0.036 −0.089 0.162 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__28DPE −0.003 −0.152 0.146 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__28DPE −0.012 −0.161 0.137 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__28DPE 0.025 −0.124 0.174 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__28DPE 0.139 −0.015 0.292 0.125 ns
Distance Temp.DPE 32°C__0DPE 28°C__42DPE 0.114 −0.016 0.243 0.161 ns
Distance Temp.DPE 32°C__14DPE 28°C__42DPE 0.057 −0.091 0.206 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__42DPE −0.001 −0.150 0.148 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__42DPE 0.045 −0.108 0.197 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__42DPE 0.034 −0.115 0.183 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__42DPE 0.061 −0.069 0.191 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__42DPE 0.021 −0.131 0.174 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__42DPE 0.012 −0.140 0.165 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__42DPE 0.049 −0.103 0.202 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__42DPE 0.163 0.006 0.320 0.034 *
Distance Temp.DPE 32°C__14DPE 32°C__0DPE −0.056 −0.182 0.069 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 32°C__0DPE −0.114 −0.240 0.011 0.115 ns
Distance Temp.DPE 32°C__28DPE 32°C__0DPE −0.069 −0.199 0.061 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__0DPE −0.080 −0.205 0.046 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 32°C__0DPE −0.053 −0.155 0.049 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__0DPE −0.092 −0.222 0.037 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__0DPE −0.101 −0.231 0.029 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__0DPE −0.064 −0.194 0.066 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__0DPE 0.049 −0.086 0.185 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 32°C__14DPE −0.058 −0.203 0.086 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 32°C__14DPE −0.013 −0.161 0.136 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__14DPE −0.024 −0.168 0.121 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 32°C__14DPE 0.003 −0.122 0.129 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__14DPE −0.036 −0.185 0.112 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__14DPE −0.045 −0.194 0.104 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__14DPE −0.008 −0.157 0.141 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__14DPE 0.105 −0.048 0.259 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 32°C__21DPE 0.046 −0.103 0.194 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__21DPE 0.035 −0.110 0.179 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 32°C__21DPE 0.061 −0.064 0.187 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__21DPE 0.022 −0.127 0.171 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__21DPE 0.013 −0.136 0.162 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__21DPE 0.050 −0.099 0.199 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__21DPE 0.164 0.010 0.317 0.025 *
Distance Temp.DPE 32°C__42DPE 32°C__28DPE −0.011 −0.160 0.138 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 32°C__28DPE 0.016 −0.114 0.146 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__28DPE −0.024 −0.176 0.129 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__28DPE −0.033 −0.185 0.120 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__28DPE 0.004 −0.148 0.157 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__28DPE 0.118 −0.039 0.275 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 32°C__42DPE 0.027 −0.099 0.152 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__42DPE −0.013 −0.161 0.136 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__42DPE −0.021 −0.170 0.127 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__42DPE 0.015 −0.133 0.164 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__42DPE 0.129 −0.024 0.283 0.209 ns
Distance Temp.DPE 35°C__14DPE 35°C__0DPE −0.039 −0.169 0.090 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 35°C__0DPE −0.048 −0.178 0.082 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__0DPE −0.011 −0.141 0.119 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__0DPE 0.102 −0.033 0.238 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 35°C__14DPE −0.009 −0.161 0.144 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__14DPE 0.028 −0.124 0.181 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__14DPE 0.142 −0.016 0.299 0.127 ns
Distance Temp.DPE 35°C__28DPE 35°C__21DPE 0.037 −0.116 0.190 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__21DPE 0.151 −0.007 0.308 0.076 ns
Distance Temp.DPE 35°C__42DPE 35°C__28DPE 0.114 −0.044 0.271 ≥0.25 ns
canberra
Distance Temp.DPE 28°C__14DPE 28°C__0DPE −0.033 −0.138 0.072 ≥0.25 ns
Distance Temp.DPE 28°C__21DPE 28°C__0DPE −0.005 −0.103 0.092 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__0DPE −0.074 −0.171 0.024 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__0DPE −0.069 −0.170 0.032 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__0DPE 0.035 −0.045 0.115 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__0DPE −0.021 −0.119 0.076 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__0DPE −0.070 −0.167 0.028 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__0DPE −0.056 −0.157 0.045 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__0DPE −0.068 −0.165 0.030 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__0DPE 0.008 −0.071 0.088 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__0DPE −0.041 −0.142 0.060 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__0DPE −0.047 −0.148 0.054 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__0DPE −0.052 −0.153 0.049 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__0DPE 0.045 −0.061 0.150 ≥0.25 ns
Distance Temp.DPE 28°C__21DPE 28°C__14DPE 0.028 −0.091 0.147 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__14DPE −0.040 −0.160 0.079 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__14DPE −0.035 −0.158 0.087 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__14DPE 0.068 −0.037 0.174 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__14DPE 0.012 −0.107 0.131 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__14DPE −0.037 −0.156 0.083 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__14DPE −0.023 −0.145 0.099 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__14DPE −0.034 −0.154 0.085 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__14DPE 0.042 −0.064 0.147 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__14DPE −0.008 −0.130 0.115 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__14DPE −0.013 −0.136 0.109 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__14DPE −0.019 −0.141 0.103 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__14DPE 0.078 −0.048 0.204 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__21DPE −0.068 −0.181 0.044 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__21DPE −0.063 −0.179 0.052 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__21DPE 0.040 −0.057 0.138 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__21DPE −0.016 −0.128 0.097 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__21DPE −0.065 −0.177 0.048 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__21DPE −0.051 −0.167 0.065 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__21DPE −0.062 −0.175 0.050 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__21DPE 0.014 −0.084 0.111 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__21DPE −0.036 −0.151 0.080 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__21DPE −0.041 −0.157 0.074 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__21DPE −0.047 −0.162 0.069 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__21DPE 0.050 −0.070 0.169 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__28DPE 0.005 −0.110 0.121 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__28DPE 0.109 0.011 0.206 0.014 *
Distance Temp.DPE 32°C__14DPE 28°C__28DPE 0.052 −0.060 0.165 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__28DPE 0.004 −0.109 0.116 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__28DPE 0.017 −0.098 0.133 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__28DPE 0.006 −0.106 0.119 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__28DPE 0.082 −0.015 0.180 0.206 ns
Distance Temp.DPE 35°C__14DPE 28°C__28DPE 0.033 −0.083 0.148 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__28DPE 0.027 −0.088 0.143 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__28DPE 0.021 −0.094 0.137 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__28DPE 0.118 −0.001 0.237 0.055 ns
Distance Temp.DPE 32°C__0DPE 28°C__42DPE 0.104 0.003 0.205 0.038 *
Distance Temp.DPE 32°C__14DPE 28°C__42DPE 0.047 −0.068 0.163 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__42DPE −0.001 −0.117 0.114 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__42DPE 0.012 −0.106 0.131 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__42DPE 0.001 −0.115 0.117 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__42DPE 0.077 −0.024 0.178 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__42DPE 0.028 −0.091 0.146 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__42DPE 0.022 −0.097 0.141 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__42DPE 0.016 −0.102 0.135 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__42DPE 0.113 −0.009 0.235 0.104 ns
Distance Temp.DPE 32°C__14DPE 32°C__0DPE −0.056 −0.154 0.041 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 32°C__0DPE −0.105 −0.202 −0.007 0.022 *
Distance Temp.DPE 32°C__28DPE 32°C__0DPE −0.091 −0.192 0.010 0.125 ns
Distance Temp.DPE 32°C__42DPE 32°C__0DPE −0.103 −0.200 −0.005 0.029 *
Distance Temp.DPE 35°C__0DPE 32°C__0DPE −0.027 −0.106 0.053 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__0DPE −0.076 −0.177 0.025 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__0DPE −0.082 −0.183 0.019 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__0DPE −0.087 −0.188 0.014 0.175 ns
Distance Temp.DPE 35°C__42DPE 32°C__0DPE 0.009 −0.096 0.115 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 32°C__14DPE −0.049 −0.161 0.064 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 32°C__14DPE −0.035 −0.151 0.081 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__14DPE −0.046 −0.159 0.066 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 32°C__14DPE 0.030 −0.068 0.127 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__14DPE −0.020 −0.135 0.096 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__14DPE −0.025 −0.141 0.090 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__14DPE −0.031 −0.147 0.085 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__14DPE 0.066 −0.054 0.185 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 32°C__21DPE 0.014 −0.102 0.129 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__21DPE 0.002 −0.110 0.115 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 32°C__21DPE 0.078 −0.019 0.176 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__21DPE 0.029 −0.087 0.145 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__21DPE 0.023 −0.092 0.139 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__21DPE 0.018 −0.098 0.133 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__21DPE 0.114 −0.005 0.234 0.076 ns
Distance Temp.DPE 32°C__42DPE 32°C__28DPE −0.011 −0.127 0.104 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 32°C__28DPE 0.065 −0.036 0.166 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__28DPE 0.015 −0.103 0.134 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__28DPE 0.010 −0.109 0.128 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__28DPE 0.004 −0.115 0.123 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__28DPE 0.101 −0.022 0.223 0.237 ns
Distance Temp.DPE 35°C__0DPE 32°C__42DPE 0.076 −0.021 0.173 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__42DPE 0.027 −0.089 0.142 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__42DPE 0.021 −0.095 0.137 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__42DPE 0.015 −0.100 0.131 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__42DPE 0.112 −0.007 0.231 0.091 ns
Distance Temp.DPE 35°C__14DPE 35°C__0DPE −0.049 −0.150 0.052 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 35°C__0DPE −0.055 −0.156 0.046 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__0DPE −0.061 −0.162 0.040 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__0DPE 0.036 −0.069 0.141 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 35°C__14DPE −0.006 −0.124 0.113 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__14DPE −0.011 −0.130 0.107 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__14DPE 0.085 −0.037 0.208 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__21DPE −0.006 −0.124 0.113 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__21DPE 0.091 −0.031 0.213 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__28DPE 0.097 −0.026 0.219 ≥0.25 ns
gunifrac
Distance Temp.DPE 28°C__14DPE 28°C__0DPE −0.005 −0.116 0.107 ≥0.25 ns
Distance Temp.DPE 28°C__21DPE 28°C__0DPE 0.038 −0.065 0.141 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__0DPE −0.010 −0.113 0.093 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__0DPE −0.033 −0.140 0.073 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__0DPE 0.067 −0.017 0.151 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__0DPE 0.018 −0.085 0.121 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__0DPE −0.023 −0.126 0.080 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__0DPE −0.014 −0.121 0.093 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__0DPE −0.026 −0.129 0.077 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__0DPE 0.029 −0.055 0.113 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__0DPE −0.015 −0.122 0.092 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__0DPE −0.020 −0.127 0.087 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__0DPE −0.065 −0.171 0.042 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__0DPE 0.091 −0.020 0.202 0.245 ns
Distance Temp.DPE 28°C__21DPE 28°C__14DPE 0.042 −0.084 0.168 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__14DPE −0.005 −0.131 0.121 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__14DPE −0.029 −0.158 0.100 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__14DPE 0.072 −0.040 0.183 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__14DPE 0.022 −0.104 0.148 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__14DPE −0.019 −0.145 0.107 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__14DPE −0.009 −0.138 0.120 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__14DPE −0.021 −0.147 0.105 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__14DPE 0.033 −0.078 0.145 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__14DPE −0.010 −0.139 0.119 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__14DPE −0.015 −0.145 0.114 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__14DPE −0.060 −0.189 0.069 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__14DPE 0.096 −0.037 0.229 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__21DPE −0.047 −0.166 0.072 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__21DPE −0.071 −0.193 0.051 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__21DPE 0.029 −0.074 0.132 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__21DPE −0.020 −0.139 0.099 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__21DPE −0.061 −0.180 0.058 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__21DPE −0.051 −0.174 0.071 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__21DPE −0.063 −0.182 0.056 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__21DPE −0.009 −0.112 0.094 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__21DPE −0.052 −0.175 0.070 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__21DPE −0.058 −0.180 0.065 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__21DPE −0.102 −0.224 0.020 0.216 ns
Distance Temp.DPE 35°C__42DPE 28°C__21DPE 0.054 −0.073 0.180 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__28DPE −0.024 −0.146 0.098 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__28DPE 0.077 −0.026 0.180 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__28DPE 0.027 −0.092 0.146 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__28DPE −0.014 −0.133 0.105 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__28DPE −0.004 −0.126 0.118 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__28DPE −0.016 −0.135 0.103 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__28DPE 0.038 −0.065 0.142 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__28DPE −0.005 −0.127 0.117 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__28DPE −0.010 −0.132 0.112 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__28DPE −0.055 −0.177 0.067 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__28DPE 0.101 −0.025 0.227 ≥0.25 ns
Distance Temp.DPE 32°C__0DPE 28°C__42DPE 0.100 −0.006 0.207 0.089 ns
Distance Temp.DPE 32°C__14DPE 28°C__42DPE 0.051 −0.071 0.173 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__42DPE 0.010 −0.112 0.132 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__42DPE 0.020 −0.106 0.145 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__42DPE 0.008 −0.114 0.130 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 28°C__42DPE 0.062 −0.045 0.169 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__42DPE 0.019 −0.107 0.144 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__42DPE 0.014 −0.112 0.139 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__42DPE −0.031 −0.157 0.094 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__42DPE 0.125 −0.005 0.254 0.071 ns
Distance Temp.DPE 32°C__14DPE 32°C__0DPE −0.049 −0.152 0.054 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 32°C__0DPE −0.090 −0.193 0.013 0.155 ns
Distance Temp.DPE 32°C__28DPE 32°C__0DPE −0.081 −0.188 0.026 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__0DPE −0.093 −0.196 0.010 0.130 ns
Distance Temp.DPE 35°C__0DPE 32°C__0DPE −0.038 −0.122 0.046 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__0DPE −0.082 −0.189 0.025 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__0DPE −0.087 −0.194 0.020 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__0DPE −0.132 −0.238 −0.025 0.003 **
Distance Temp.DPE 35°C__42DPE 32°C__0DPE 0.024 −0.087 0.135 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 32°C__14DPE −0.041 −0.160 0.078 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 32°C__14DPE −0.031 −0.154 0.091 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__14DPE −0.043 −0.162 0.076 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 32°C__14DPE 0.011 −0.092 0.114 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__14DPE −0.032 −0.155 0.090 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__14DPE −0.037 −0.160 0.085 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__14DPE −0.082 −0.204 0.040 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__14DPE 0.074 −0.053 0.200 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 32°C__21DPE 0.010 −0.113 0.132 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__21DPE −0.002 −0.121 0.117 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 32°C__21DPE 0.052 −0.051 0.155 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__21DPE 0.009 −0.114 0.131 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__21DPE 0.004 −0.119 0.126 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__21DPE −0.041 −0.163 0.081 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__21DPE 0.115 −0.011 0.241 0.119 ns
Distance Temp.DPE 32°C__42DPE 32°C__28DPE −0.012 −0.134 0.110 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 32°C__28DPE 0.043 −0.064 0.149 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__28DPE −0.001 −0.126 0.124 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__28DPE −0.006 −0.132 0.119 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__28DPE −0.051 −0.176 0.075 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__28DPE 0.105 −0.024 0.234 ≥0.25 ns
Distance Temp.DPE 35°C__0DPE 32°C__42DPE 0.054 −0.049 0.157 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__42DPE 0.011 −0.111 0.133 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__42DPE 0.006 −0.117 0.128 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__42DPE −0.039 −0.161 0.083 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__42DPE 0.117 −0.009 0.243 0.102 ns
Distance Temp.DPE 35°C__14DPE 35°C__0DPE −0.044 −0.150 0.063 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 35°C__0DPE −0.049 −0.155 0.058 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__0DPE −0.093 −0.200 0.013 0.160 ns
Distance Temp.DPE 35°C__42DPE 35°C__0DPE 0.062 −0.049 0.174 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 35°C__14DPE −0.005 −0.130 0.120 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__14DPE −0.050 −0.175 0.076 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__14DPE 0.106 −0.023 0.235 0.244 ns
Distance Temp.DPE 35°C__28DPE 35°C__21DPE −0.045 −0.170 0.081 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__21DPE 0.111 −0.018 0.240 0.180 ns
Distance Temp.DPE 35°C__42DPE 35°C__28DPE 0.156 0.027 0.285 0.005 **

3) Parasitic worm burden differs by water temperature

Fig. 3 Histological sections stained with ### stain in zebrafish exposed to Pseudocapillaria tomentosa examined at ### days post exposure. (A) Nematodes in intestinal lumen. Arrow = ###. (B) Infection outcome analysis of fish exposed to P. tomentosa (n = 89) by temperature. Fish reared at 28°C and 32°C water temperatures had significantly different infection burden to fish reared at 35°C water temperature. Only one fish reared at 35°C was identified as being positively infected by wet mount.

*3A)

3B)

4) Water temperature differentially stratifies gut microbiome communities of parasite exposed fish

Fig. 4 Effects of parasite exposure and water temperature on gut microbiomes of zebrafish. (A) Simpson’s Index of diversity shows that gut microbiome diversity significantly differs between fish reared at 28°C water temperature to fish reared at 32°C and 35°C water temperatures. (B) Capscale ordination based on the Bray-Curtis dissimilarity of gut microbiome composition. The analysis shows that gut microbiome composition significantly differs between parasite exposed fish reared at different water temperatures. (C) Simpson’s Index of diversity shows microbial gut diversity decreases with time from 0 days post exposure (dpe) to 42 dpe in parasite exposed fish reared at 28°C water temperature. (D) Capscale ordination of gut microbiome composition based on the Canberra dissimilarity constrained on the main effects of water temperature and time, and their interaction. The analysis shows that shows that gut microbiome composition differs between parasite exposed fish across time depending on water temperature. Ribbons and ellipses indicate 95% confidence interval. Only statistically significant relationships are shown. A “*” indicates statistical significance below the “0.05” level. Black arrows indicate direction of greatest change in the indicated covariates.

4A)

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GLM
GLM Results
glm(Alpha.Score ~ Temperature); Exposed fish
term estimate std.error statistic p.value p.adj.sig
Shannon
(Intercept) −0.936 0.233 −4.021 <0.001 ***
Temperature32 0.909 0.301 3.017 0.003 **
Temperature35 0.516 0.296 1.741 0.085 ns
Simpson
(Intercept) −1.002 0.243 −4.130 <0.001 ****
Temperature32 1.057 0.312 3.384 0.001 **
Temperature35 0.772 0.306 2.525 0.013 *
Richness
(Intercept) −0.681 0.190 −3.583 <0.001 ***
Temperature32 0.703 0.251 2.800 0.006 **
Temperature35 0.242 0.247 0.978 ≥0.25 ns
ANOVA
ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature), type = 2); Exposed fish
term statistic df p.value sig
Shannon
Temperature 9.440 2.000 0.009 **
Simpson
Temperature 12.497 2.000 0.002 **
Richness
Temperature 8.575 2.000 0.014 *
Tukey
Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature); Exposed fish
term .y. group1 group2 estimate std.error statistic adj.p.value Variable
Shannon
Temperature Alpha.Score 32 28 0.909 0.301 3.017 0.007 Temperature
Temperature Alpha.Score 35 28 0.516 0.296 1.741 0.190 Temperature
Temperature Alpha.Score 35 32 −0.393 0.265 −1.482 ≥0.25 Temperature
Simpson
Temperature Alpha.Score 32 28 1.057 0.312 3.384 0.002 Temperature
Temperature Alpha.Score 35 28 0.772 0.306 2.525 0.031 Temperature
Temperature Alpha.Score 35 32 −0.285 0.270 −1.054 ≥0.25 Temperature
Richness
Temperature Alpha.Score 32 28 0.703 0.251 2.800 0.014 Temperature
Temperature Alpha.Score 35 28 0.242 0.247 0.978 ≥0.25 Temperature
Temperature Alpha.Score 35 32 −0.461 0.228 −2.028 0.105 Temperature

4B)

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Capscale
ADONIS2
adonis2(Beta Distance ~ Temperature); Exposed fish
term df SumOfSqs R2 statistic p.value sig
bray
Temperature 2.000 5.218 0.251 14.447 0.001 ***
Residual 86.000 15.530 0.749 NA NA NA
Total 88.000 20.748 1.000 NA NA NA
canberra
Temperature 2.000 3.573 0.117 5.701 0.001 ***
Residual 86.000 26.947 0.883 NA NA NA
Total 88.000 30.520 1.000 NA NA NA
gunifrac
Temperature 2.000 3.363 0.212 11.590 0.001 ***
Residual 86.000 12.478 0.788 NA NA NA
Total 88.000 15.841 1.000 NA NA NA
Dispersion (ANOVA)
ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature); Exposed fish
term df sumsq meansq statistic p.value sig
bray
Temperature 2.000 0.364 0.182 8.708 <0.001 ***
Residual 86.000 1.796 0.021 NA NA NA
canberra
Temperature 2.000 0.314 0.157 17.419 <0.001 ****
Residual 86.000 0.774 0.009 NA NA NA
gunifrac
Temperature 2.000 0.310 0.155 13.567 <0.001 ****
Residual 86.000 0.983 0.011 NA NA NA
Dispersion (Tukey)
Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature); Exposed fish
.y. term group1 group2 estimate conf.low conf.high adj.p.value sig
bray
Distance Temperature 32 28 −0.043 −0.136 0.051 ≥0.25 ns
Distance Temperature 35 28 −0.150 −0.241 −0.059 <0.001 ***
Distance Temperature 35 32 −0.108 −0.194 −0.021 0.011 *
canberra
Distance Temperature 32 28 −0.110 −0.171 −0.049 <0.001 ***
Distance Temperature 35 28 −0.144 −0.204 −0.085 <0.001 ****
Distance Temperature 35 32 −0.034 −0.091 0.022 ≥0.25 ns
gunifrac
Distance Temperature 32 28 −0.056 −0.125 0.013 0.133 ns
Distance Temperature 35 28 −0.143 −0.211 −0.076 <0.001 ****
Distance Temperature 35 32 −0.087 −0.151 −0.023 0.005 **

4C)

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GLM
GLM Results
glm(Alpha.Score ~ Temperature*DPE); Exposed fish
term estimate std.error statistic p.value p.adj.sig
Shannon
(Intercept) −0.134 0.594 −0.226 ≥0.25 ns
Temperature32 0.228 0.774 0.295 ≥0.25 ns
Temperature35 −0.428 0.777 −0.551 ≥0.25 ns
DPE −0.031 0.022 −1.425 0.158 ns
Temperature32:DPE 0.026 0.028 0.917 ≥0.25 ns
Temperature35:DPE 0.036 0.028 1.300 0.197 ns
Simpson
(Intercept) −0.282 0.617 −0.456 ≥0.25 ns
Temperature32 0.336 0.801 0.420 ≥0.25 ns
Temperature35 −0.172 0.799 −0.215 ≥0.25 ns
DPE −0.028 0.022 −1.234 0.221 ns
Temperature32:DPE 0.028 0.029 0.946 ≥0.25 ns
Temperature35:DPE 0.036 0.029 1.261 0.211 ns
Richness
(Intercept) 0.316 0.480 0.657 ≥0.25 ns
Temperature32 −0.034 0.636 −0.054 ≥0.25 ns
Temperature35 −0.596 0.638 −0.934 ≥0.25 ns
DPE −0.038 0.017 −2.191 0.031 *
Temperature32:DPE 0.028 0.023 1.199 0.234 ns
Temperature35:DPE 0.032 0.023 1.405 0.164 ns
ANOVA
ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature*Time), type = 2); Exposed fish
term statistic df p.value sig
Shannon
Temperature 8.789 2.000 0.012 *
DPE 0.523 1.000 ≥0.25 ns
Temperature:DPE 1.759 2.000 ≥0.25 ns
Simpson
Temperature 11.871 2.000 0.003 **
DPE 0.120 1.000 ≥0.25 ns
Temperature:DPE 1.684 2.000 ≥0.25 ns
Richness
Temperature 7.911 2.000 0.019 *
DPE 3.435 1.000 0.064 ns
Temperature:DPE 2.255 2.000 ≥0.25 ns
Tukey
Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature*Time); Exposed fish
Temperature .y. term group1 group2 estimate std.error statistic adj.p.value Variable Group
Shannon
28 Alpha.Score DPE 21 14 −3.311 1.822 −1.817 0.246 DPE 28
28 Alpha.Score DPE 28 14 −2.027 1.023 −1.982 0.178 DPE 28
28 Alpha.Score DPE 42 14 −1.024 0.705 −1.452 ≥0.25 DPE 28
28 Alpha.Score DPE 28 21 1.284 1.982 0.648 ≥0.25 DPE 28
28 Alpha.Score DPE 42 21 2.287 1.838 1.245 ≥0.25 DPE 28
28 Alpha.Score DPE 42 28 1.004 1.050 0.956 ≥0.25 DPE 28
32 Alpha.Score DPE 21 14 −0.536 0.537 −0.999 ≥0.25 DPE 32
32 Alpha.Score DPE 28 14 0.056 0.558 0.099 ≥0.25 DPE 32
32 Alpha.Score DPE 42 14 −0.271 0.532 −0.509 ≥0.25 DPE 32
32 Alpha.Score DPE 28 21 0.592 0.606 0.977 ≥0.25 DPE 32
32 Alpha.Score DPE 42 21 0.266 0.581 0.457 ≥0.25 DPE 32
32 Alpha.Score DPE 42 28 −0.326 0.601 −0.543 ≥0.25 DPE 32
35 Alpha.Score DPE 21 14 −0.246 0.270 −0.909 ≥0.25 DPE 35
35 Alpha.Score DPE 28 14 −0.292 0.271 −1.075 ≥0.25 DPE 35
35 Alpha.Score DPE 42 14 0.097 0.258 0.377 ≥0.25 DPE 35
35 Alpha.Score DPE 28 21 −0.046 0.283 −0.162 ≥0.25 DPE 35
35 Alpha.Score DPE 42 21 0.343 0.270 1.273 ≥0.25 DPE 35
35 Alpha.Score DPE 42 28 0.389 0.271 1.438 ≥0.25 DPE 35
Simpson
28 Alpha.Score DPE 21 14 −3.141 1.820 −1.725 ≥0.25 DPE 28
28 Alpha.Score DPE 28 14 −1.994 1.072 −1.861 0.226 DPE 28
28 Alpha.Score DPE 42 14 −0.925 0.721 −1.283 ≥0.25 DPE 28
28 Alpha.Score DPE 28 21 1.147 2.001 0.573 ≥0.25 DPE 28
28 Alpha.Score DPE 42 21 2.215 1.837 1.206 ≥0.25 DPE 28
28 Alpha.Score DPE 42 28 1.069 1.100 0.971 ≥0.25 DPE 28
32 Alpha.Score DPE 21 14 −0.416 0.550 −0.756 ≥0.25 DPE 32
32 Alpha.Score DPE 28 14 0.275 0.580 0.474 ≥0.25 DPE 32
32 Alpha.Score DPE 42 14 −0.138 0.546 −0.252 ≥0.25 DPE 32
32 Alpha.Score DPE 28 21 0.691 0.626 1.103 ≥0.25 DPE 32
32 Alpha.Score DPE 42 21 0.278 0.595 0.467 ≥0.25 DPE 32
32 Alpha.Score DPE 42 28 −0.413 0.623 −0.662 ≥0.25 DPE 32
35 Alpha.Score DPE 21 14 −0.374 0.297 −1.258 ≥0.25 DPE 35
35 Alpha.Score DPE 28 14 −0.285 0.296 −0.966 ≥0.25 DPE 35
35 Alpha.Score DPE 42 14 0.142 0.284 0.502 ≥0.25 DPE 35
35 Alpha.Score DPE 28 21 0.088 0.308 0.287 ≥0.25 DPE 35
35 Alpha.Score DPE 42 21 0.516 0.297 1.738 ≥0.25 DPE 35
35 Alpha.Score DPE 42 28 0.428 0.295 1.448 ≥0.25 DPE 35
Richness
28 Alpha.Score DPE 21 14 −2.219 1.016 −2.185 0.123 DPE 28
28 Alpha.Score DPE 28 14 −1.585 0.795 −1.993 0.184 DPE 28
28 Alpha.Score DPE 42 14 −1.239 0.650 −1.907 0.218 DPE 28
28 Alpha.Score DPE 28 21 0.634 1.133 0.560 ≥0.25 DPE 28
28 Alpha.Score DPE 42 21 0.980 1.036 0.946 ≥0.25 DPE 28
28 Alpha.Score DPE 42 28 0.346 0.821 0.421 ≥0.25 DPE 28
32 Alpha.Score DPE 21 14 −0.561 0.412 −1.363 ≥0.25 DPE 32
32 Alpha.Score DPE 28 14 −0.470 0.430 −1.091 ≥0.25 DPE 32
32 Alpha.Score DPE 42 14 −0.357 0.410 −0.871 ≥0.25 DPE 32
32 Alpha.Score DPE 28 21 0.092 0.462 0.199 ≥0.25 DPE 32
32 Alpha.Score DPE 42 21 0.204 0.444 0.460 ≥0.25 DPE 32
32 Alpha.Score DPE 42 28 0.112 0.461 0.243 ≥0.25 DPE 32
35 Alpha.Score DPE 21 14 −0.338 0.242 −1.396 ≥0.25 DPE 35
35 Alpha.Score DPE 28 14 −0.329 0.242 −1.361 ≥0.25 DPE 35
35 Alpha.Score DPE 42 14 −0.233 0.233 −1.000 ≥0.25 DPE 35
35 Alpha.Score DPE 28 21 0.009 0.252 0.035 ≥0.25 DPE 35
35 Alpha.Score DPE 42 21 0.105 0.244 0.430 ≥0.25 DPE 35
35 Alpha.Score DPE 42 28 0.096 0.244 0.394 ≥0.25 DPE 35

4D)

Tables
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Capscale
ADONIS2
adonis2(Beta Distance ~ Temperature*DPE); Exposed fish
term df SumOfSqs R2 statistic p.value sig
bray
Temperature 2.000 5.218 0.251 14.843 0.001 ***
DPE 1.000 0.394 0.019 2.241 0.040 *
Temperature:DPE 2.000 0.548 0.026 1.558 0.110 ns
Residual 83.000 14.589 0.703 NA NA NA
Total 88.000 20.748 1.000 NA NA NA
canberra
Temperature 2.000 3.573 0.117 5.847 0.001 ***
DPE 1.000 0.674 0.022 2.207 0.004 **
Temperature:DPE 2.000 0.913 0.030 1.494 0.015 *
Residual 83.000 25.360 0.831 NA NA NA
Total 88.000 30.520 1.000 NA NA NA
gunifrac
Temperature 2.000 3.363 0.212 11.847 0.001 ***
DPE 1.000 0.261 0.016 1.837 0.102 ns
Temperature:DPE 2.000 0.436 0.028 1.535 0.134 ns
Residual 83.000 11.781 0.744 NA NA NA
Total 88.000 15.841 1.000 NA NA NA
Dispersion (ANOVA)
ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature); Exposed fish
term df sumsq meansq statistic p.value sig
bray
Temp.DPE 11.000 0.624 0.057 3.261 0.001 **
Residual 77.000 1.339 0.017 NA NA NA
canberra
Temp.DPE 11.000 0.316 0.029 2.748 0.005 **
Residual 77.000 0.805 0.010 NA NA NA
gunifrac
Temp.DPE 11.000 0.420 0.038 3.232 0.001 **
Residual 77.000 0.909 0.012 NA NA NA
Dispersion (Tukey)
Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature*DPE); Exposed fish
.y. term group1 group2 estimate conf.low conf.high adj.p.value sig
bray
Distance Temp.DPE 28°C__21DPE 28°C__14DPE −0.213 −0.485 0.059 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__14DPE −0.258 −0.512 −0.005 0.042 *
Distance Temp.DPE 28°C__42DPE 28°C__14DPE −0.063 −0.285 0.160 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__14DPE −0.059 −0.270 0.152 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__14DPE −0.089 −0.319 0.141 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__14DPE −0.123 −0.363 0.117 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__14DPE −0.142 −0.372 0.088 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__14DPE −0.202 −0.418 0.014 0.090 ns
Distance Temp.DPE 35°C__21DPE 28°C__14DPE −0.170 −0.393 0.052 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__14DPE −0.289 −0.511 −0.067 0.002 **
Distance Temp.DPE 35°C__42DPE 28°C__14DPE −0.192 −0.408 0.024 0.130 ns
Distance Temp.DPE 28°C__28DPE 28°C__21DPE −0.046 −0.344 0.253 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__21DPE 0.150 −0.122 0.422 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__21DPE 0.153 −0.110 0.416 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__21DPE 0.124 −0.154 0.403 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__21DPE 0.090 −0.197 0.377 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__21DPE 0.071 −0.208 0.350 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__21DPE 0.011 −0.256 0.278 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__21DPE 0.042 −0.230 0.315 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__21DPE −0.077 −0.349 0.196 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__21DPE 0.021 −0.246 0.288 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__28DPE 0.196 −0.058 0.449 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__28DPE 0.199 −0.044 0.443 0.220 ns
Distance Temp.DPE 32°C__21DPE 28°C__28DPE 0.170 −0.091 0.430 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__28DPE 0.136 −0.133 0.405 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__28DPE 0.117 −0.144 0.377 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__28DPE 0.057 −0.191 0.305 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__28DPE 0.088 −0.165 0.341 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__28DPE −0.031 −0.284 0.222 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__28DPE 0.066 −0.182 0.314 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__42DPE 0.003 −0.207 0.214 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__42DPE −0.026 −0.256 0.204 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__42DPE −0.060 −0.300 0.180 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__42DPE −0.079 −0.309 0.151 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__42DPE −0.139 −0.355 0.077 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__42DPE −0.108 −0.330 0.115 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__42DPE −0.227 −0.449 −0.004 0.042 *
Distance Temp.DPE 35°C__42DPE 28°C__42DPE −0.129 −0.345 0.087 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 32°C__14DPE −0.029 −0.248 0.190 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 32°C__14DPE −0.063 −0.293 0.166 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__14DPE −0.082 −0.301 0.137 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__14DPE −0.142 −0.347 0.062 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__14DPE −0.111 −0.322 0.100 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__14DPE −0.230 −0.441 −0.019 0.021 *
Distance Temp.DPE 35°C__42DPE 32°C__14DPE −0.133 −0.337 0.072 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 32°C__21DPE −0.034 −0.281 0.213 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__21DPE −0.053 −0.291 0.185 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__21DPE −0.113 −0.337 0.111 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__21DPE −0.082 −0.312 0.148 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__21DPE −0.201 −0.431 0.029 0.148 ns
Distance Temp.DPE 35°C__42DPE 32°C__21DPE −0.103 −0.327 0.121 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__28DPE −0.019 −0.266 0.228 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__28DPE −0.079 −0.313 0.155 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__28DPE −0.048 −0.288 0.192 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__28DPE −0.167 −0.407 0.073 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__28DPE −0.069 −0.304 0.165 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__42DPE −0.060 −0.284 0.164 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__42DPE −0.029 −0.259 0.201 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__42DPE −0.148 −0.378 0.082 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__42DPE −0.050 −0.274 0.174 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 35°C__14DPE 0.031 −0.185 0.247 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__14DPE −0.088 −0.304 0.128 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__14DPE 0.010 −0.200 0.219 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__21DPE −0.119 −0.341 0.103 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__21DPE −0.022 −0.238 0.194 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__28DPE 0.097 −0.119 0.313 ≥0.25 ns
canberra
Distance Temp.DPE 28°C__21DPE 28°C__14DPE −0.004 −0.215 0.207 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__14DPE 0.040 −0.156 0.237 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__14DPE 0.059 −0.113 0.232 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__14DPE −0.043 −0.206 0.121 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__14DPE −0.044 −0.223 0.134 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__14DPE −0.102 −0.288 0.084 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__14DPE −0.084 −0.262 0.095 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__14DPE −0.097 −0.265 0.070 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__14DPE −0.097 −0.269 0.075 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__14DPE −0.142 −0.314 0.031 0.215 ns
Distance Temp.DPE 35°C__42DPE 28°C__14DPE −0.103 −0.271 0.064 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__21DPE 0.044 −0.187 0.276 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__21DPE 0.064 −0.148 0.275 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__21DPE −0.039 −0.243 0.165 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__21DPE −0.040 −0.256 0.176 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__21DPE −0.097 −0.320 0.125 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__21DPE −0.079 −0.295 0.137 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__21DPE −0.093 −0.300 0.114 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__21DPE −0.093 −0.304 0.118 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__21DPE −0.137 −0.348 0.074 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__21DPE −0.099 −0.306 0.108 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__28DPE 0.019 −0.177 0.216 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__28DPE −0.083 −0.272 0.106 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__28DPE −0.085 −0.286 0.117 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__28DPE −0.142 −0.351 0.067 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__28DPE −0.124 −0.326 0.078 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__28DPE −0.137 −0.329 0.055 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__28DPE −0.137 −0.334 0.059 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__28DPE −0.182 −0.378 0.015 0.097 ns
Distance Temp.DPE 35°C__42DPE 28°C__28DPE −0.144 −0.336 0.049 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__42DPE −0.102 −0.266 0.061 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__42DPE −0.104 −0.282 0.075 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__42DPE −0.161 −0.347 0.025 0.157 ns
Distance Temp.DPE 32°C__42DPE 28°C__42DPE −0.143 −0.321 0.036 0.247 ns
Distance Temp.DPE 35°C__14DPE 28°C__42DPE −0.156 −0.324 0.011 0.091 ns
Distance Temp.DPE 35°C__21DPE 28°C__42DPE −0.156 −0.329 0.016 0.112 ns
Distance Temp.DPE 35°C__28DPE 28°C__42DPE −0.201 −0.373 −0.028 0.010 **
Distance Temp.DPE 35°C__42DPE 28°C__42DPE −0.163 −0.330 0.005 0.065 ns
Distance Temp.DPE 32°C__21DPE 32°C__14DPE −0.001 −0.171 0.168 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 32°C__14DPE −0.059 −0.237 0.119 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__14DPE −0.041 −0.211 0.129 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__14DPE −0.054 −0.212 0.104 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__14DPE −0.054 −0.218 0.109 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__14DPE −0.099 −0.262 0.065 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__14DPE −0.060 −0.219 0.098 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 32°C__21DPE −0.057 −0.249 0.134 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__21DPE −0.039 −0.224 0.145 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__21DPE −0.053 −0.226 0.121 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__21DPE −0.053 −0.231 0.126 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__21DPE −0.097 −0.276 0.081 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__21DPE −0.059 −0.233 0.115 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__28DPE 0.018 −0.174 0.210 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__28DPE 0.005 −0.177 0.186 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__28DPE 0.005 −0.182 0.191 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__28DPE −0.040 −0.226 0.146 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__28DPE −0.002 −0.183 0.180 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__42DPE −0.013 −0.187 0.160 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__42DPE −0.014 −0.192 0.165 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__42DPE −0.058 −0.236 0.120 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__42DPE −0.020 −0.194 0.154 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 35°C__14DPE 0.000 −0.168 0.167 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__14DPE −0.044 −0.212 0.123 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__14DPE −0.006 −0.169 0.156 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__21DPE −0.044 −0.217 0.128 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__21DPE −0.006 −0.174 0.161 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__28DPE 0.038 −0.129 0.206 ≥0.25 ns
gunifrac
Distance Temp.DPE 28°C__21DPE 28°C__14DPE −0.191 −0.415 0.033 0.173 ns
Distance Temp.DPE 28°C__28DPE 28°C__14DPE −0.061 −0.270 0.147 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__14DPE 0.018 −0.165 0.201 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__14DPE −0.024 −0.197 0.150 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__14DPE −0.027 −0.216 0.163 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__14DPE −0.100 −0.298 0.098 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__14DPE −0.069 −0.259 0.120 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__14DPE −0.131 −0.309 0.047 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__14DPE −0.137 −0.320 0.046 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__14DPE −0.198 −0.382 −0.015 0.022 *
Distance Temp.DPE 35°C__42DPE 28°C__14DPE −0.134 −0.312 0.044 ≥0.25 ns
Distance Temp.DPE 28°C__28DPE 28°C__21DPE 0.130 −0.116 0.375 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__21DPE 0.209 −0.015 0.433 0.092 ns
Distance Temp.DPE 32°C__14DPE 28°C__21DPE 0.167 −0.049 0.384 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__21DPE 0.164 −0.065 0.394 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__21DPE 0.091 −0.146 0.327 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__21DPE 0.122 −0.108 0.351 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__21DPE 0.060 −0.161 0.280 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__21DPE 0.054 −0.170 0.278 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__21DPE −0.007 −0.232 0.217 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__21DPE 0.057 −0.163 0.277 ≥0.25 ns
Distance Temp.DPE 28°C__42DPE 28°C__28DPE 0.079 −0.130 0.288 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__28DPE 0.038 −0.163 0.238 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__28DPE 0.035 −0.180 0.249 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__28DPE −0.039 −0.260 0.183 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__28DPE −0.008 −0.222 0.206 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__28DPE −0.070 −0.274 0.134 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 28°C__28DPE −0.075 −0.284 0.133 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 28°C__28DPE −0.137 −0.346 0.072 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 28°C__28DPE −0.072 −0.277 0.132 ≥0.25 ns
Distance Temp.DPE 32°C__14DPE 28°C__42DPE −0.041 −0.215 0.132 ≥0.25 ns
Distance Temp.DPE 32°C__21DPE 28°C__42DPE −0.045 −0.234 0.145 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 28°C__42DPE −0.118 −0.316 0.080 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 28°C__42DPE −0.087 −0.277 0.102 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 28°C__42DPE −0.149 −0.327 0.029 0.190 ns
Distance Temp.DPE 35°C__21DPE 28°C__42DPE −0.155 −0.338 0.029 0.182 ns
Distance Temp.DPE 35°C__28DPE 28°C__42DPE −0.216 −0.399 −0.033 0.008 **
Distance Temp.DPE 35°C__42DPE 28°C__42DPE −0.151 −0.329 0.026 0.173 ns
Distance Temp.DPE 32°C__21DPE 32°C__14DPE −0.003 −0.184 0.177 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 32°C__14DPE −0.076 −0.266 0.113 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__14DPE −0.046 −0.226 0.135 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__14DPE −0.108 −0.276 0.060 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__14DPE −0.113 −0.287 0.061 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__14DPE −0.175 −0.348 −0.001 0.047 *
Distance Temp.DPE 35°C__42DPE 32°C__14DPE −0.110 −0.278 0.058 ≥0.25 ns
Distance Temp.DPE 32°C__28DPE 32°C__21DPE −0.073 −0.277 0.130 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__21DPE −0.043 −0.238 0.153 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__21DPE −0.105 −0.289 0.080 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__21DPE −0.110 −0.300 0.079 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__21DPE −0.172 −0.361 0.018 0.113 ns
Distance Temp.DPE 35°C__42DPE 32°C__21DPE −0.107 −0.292 0.078 ≥0.25 ns
Distance Temp.DPE 32°C__42DPE 32°C__28DPE 0.031 −0.173 0.234 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__28DPE −0.031 −0.224 0.162 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__28DPE −0.037 −0.234 0.161 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__28DPE −0.098 −0.296 0.099 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__28DPE −0.034 −0.227 0.159 ≥0.25 ns
Distance Temp.DPE 35°C__14DPE 32°C__42DPE −0.062 −0.247 0.122 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 32°C__42DPE −0.067 −0.257 0.122 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 32°C__42DPE −0.129 −0.319 0.060 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 32°C__42DPE −0.064 −0.249 0.120 ≥0.25 ns
Distance Temp.DPE 35°C__21DPE 35°C__14DPE −0.005 −0.183 0.173 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__14DPE −0.067 −0.245 0.111 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__14DPE −0.002 −0.175 0.170 ≥0.25 ns
Distance Temp.DPE 35°C__28DPE 35°C__21DPE −0.062 −0.245 0.121 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__21DPE 0.003 −0.175 0.181 ≥0.25 ns
Distance Temp.DPE 35°C__42DPE 35°C__28DPE 0.065 −0.113 0.243 ≥0.25 ns

5) Water temperature influences gut microbiome’s sensitivity to parasite exposure

Fig. 5 Water temperature influences gut microbiome response to Pseudocapillaria tomentosa. (A) Simpson’s Index for diversity of parasite unexposed and pre-exposed fish at 0 days post exposure (dpe). Prior to parasite exposure gut microbial alpha-diversity does not significantly differ between fish reared at the same water temperature. (B) Simpson’s Index for diversity of parasite unexposed and exposed fish. Gut microbial alpha-diversity significantly differs between parasite exposed fish reared at 28°C and 32°C water temperature relative to unexposed control fish, but gut microbial alpha-diversity does not differ between parasite unexposed and exposed fish reared at 35°C water temperature. Capscale ordinations based on the Bray-Curtis dissimilarity of gut microbiome composition constrained on the main and interaction effects of temperature and parasite exposure (treatment) (C) of pre-exposed samples at 0 dpe, and (D) post-exposed samples after 0 dpe. The analysis shows that gut microbiome composition of fish significantly differs between water temperature and parasite exposure in pre- and post-exposed fish. However, the interaction effect between temperature and parasite exposure is not significant in pre-exposed fish, but is significant in post-exposed fish. Ribbons and ellipses indicate 95% confidence interval. Ribbons and ellipses indicate 95% confidence interval. Only statistically significant relationships are shown. A “*” indicates statistical significance below the “0.05” level. Black arrows indicate statistically significant covariates and direction of greatest change in the indicated covariates.

5A)

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GLM
GLM Results
glm(Alpha.Score ~ Temperature*Treatment); Pre-Exposed fish
term estimate std.error statistic p.value p.adj.sig
Shannon
(Intercept) −0.218 0.269 −0.811 ≥0.25 ns
Temperature32 0.345 0.379 0.910 ≥0.25 ns
Temperature35 0.693 0.384 1.803 0.077 ns
TreatmentExposed 0.274 0.379 0.723 ≥0.25 ns
Temperature32:TreatmentExposed 0.311 0.544 0.572 ≥0.25 ns
Temperature35:TreatmentExposed 0.041 0.550 0.075 ≥0.25 ns
Simpson
(Intercept) −0.241 0.252 −0.958 ≥0.25 ns
Temperature32 0.352 0.355 0.990 ≥0.25 ns
Temperature35 0.961 0.367 2.619 0.011 *
TreatmentExposed 0.223 0.355 0.627 ≥0.25 ns
Temperature32:TreatmentExposed 0.486 0.513 0.948 ≥0.25 ns
Temperature35:TreatmentExposed 0.017 0.525 0.033 ≥0.25 ns
Richness
(Intercept) −0.145 0.270 −0.538 ≥0.25 ns
Temperature32 0.550 0.385 1.430 0.158 ns
Temperature35 0.043 0.381 0.112 ≥0.25 ns
TreatmentExposed 0.402 0.382 1.050 ≥0.25 ns
Temperature32:TreatmentExposed 0.020 0.554 0.036 ≥0.25 ns
Temperature35:TreatmentExposed 0.287 0.545 0.526 ≥0.25 ns
ANOVA
ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature*Treatment), type = 2); Pre-Exposed fish
term statistic df p.value sig
Shannon
Temperature 7.264 2.000 0.026 *
Treatment 3.044 1.000 0.081 ns
Temperature:Treatment 0.380 2.000 ≥0.25 ns
Simpson
Temperature 14.462 2.000 <0.001 ***
Treatment 3.406 1.000 0.065 ns
Temperature:Treatment 1.122 2.000 ≥0.25 ns
Richness
Temperature 4.311 2.000 0.116 ns
Treatment 5.067 1.000 0.024 *
Temperature:Treatment 0.340 2.000 ≥0.25 ns
Tukey
Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature*Treatment); Pre-Exposed fish
Temperature .y. term group1 group2 estimate std.error statistic adj.p.value Variable Group
Shannon
28 Alpha.Score Treatment Exposed Control 0.274 0.364 0.753 ≥0.25 Treatment 28
32 Alpha.Score Treatment Exposed Control 0.585 0.451 1.298 0.194 Treatment 32
35 Alpha.Score Treatment Exposed Control 0.315 0.344 0.917 ≥0.25 Treatment 35
Simpson
28 Alpha.Score Treatment Exposed Control 0.223 0.337 0.662 ≥0.25 Treatment 28
32 Alpha.Score Treatment Exposed Control 0.709 0.426 1.663 0.096 Treatment 32
35 Alpha.Score Treatment Exposed Control 0.240 0.339 0.708 ≥0.25 Treatment 35
Richness
28 Alpha.Score Treatment Exposed Control 0.402 0.361 1.112 ≥0.25 Treatment 28
32 Alpha.Score Treatment Exposed Control 0.422 0.411 1.027 ≥0.25 Treatment 32
35 Alpha.Score Treatment Exposed Control 0.689 0.400 1.721 0.085 Treatment 35

5B)

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GLM
GLM Results
glm(Alpha.Score ~ Temperature*Treatment); Post-Exposed fish
term estimate std.error statistic p.value p.adj.sig
Shannon
(Intercept) −0.441 0.126 −3.499 <0.001 ***
Temperature32 −0.064 0.177 −0.364 ≥0.25 ns
Temperature35 −0.087 0.182 −0.476 ≥0.25 ns
TreatmentExposed −0.539 0.210 −2.562 0.011 *
Temperature32:TreatmentExposed 0.953 0.280 3.405 <0.001 ***
Temperature35:TreatmentExposed 0.561 0.281 1.996 0.047 *
Simpson
(Intercept) −0.299 0.133 −2.252 0.025 *
Temperature32 −0.059 0.186 −0.319 ≥0.25 ns
Temperature35 0.080 0.190 0.419 ≥0.25 ns
TreatmentExposed −0.724 0.225 −3.223 0.001 **
Temperature32:TreatmentExposed 1.106 0.298 3.715 <0.001 ***
Temperature35:TreatmentExposed 0.673 0.297 2.270 0.024 *
Richness
(Intercept) −0.206 0.101 −2.040 0.043 *
Temperature32 −0.048 0.141 −0.343 ≥0.25 ns
Temperature35 −0.317 0.147 −2.164 0.032 *
TreatmentExposed −0.228 0.160 −1.421 0.157 ns
Temperature32:TreatmentExposed 0.829 0.219 3.776 <0.001 ***
Temperature35:TreatmentExposed 0.691 0.219 3.155 0.002 **
ANOVA
ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature*Treatment), type = 2); Post-Exposed fish
term statistic df p.value sig
Shannon
Temperature 5.908 2.000 0.052 ns
Treatment 0.001 1.000 ≥0.25 ns
Temperature:Treatment 11.784 2.000 0.003 **
Simpson
Temperature 8.400 2.000 0.015 *
Treatment 0.547 1.000 ≥0.25 ns
Temperature:Treatment 14.128 2.000 <0.001 ***
Richness
Temperature 11.325 2.000 0.003 **
Treatment 11.824 1.000 <0.001 ***
Temperature:Treatment 16.178 2.000 <0.001 ***
Tukey
Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature*Treatment); Post-Exposed fish
Temperature .y. term group1 group2 estimate std.error statistic adj.p.value Variable Group
Shannon
28 Alpha.Score Treatment Exposed Control −0.539 0.280 −1.924 0.054 Treatment 28
32 Alpha.Score Treatment Exposed Control 0.414 0.180 2.298 0.022 Treatment 32
35 Alpha.Score Treatment Exposed Control 0.022 0.111 0.200 ≥0.25 Treatment 35
Simpson
28 Alpha.Score Treatment Exposed Control −0.724 0.290 −2.492 0.013 Treatment 28
32 Alpha.Score Treatment Exposed Control 0.382 0.190 2.013 0.044 Treatment 32
35 Alpha.Score Treatment Exposed Control −0.051 0.130 −0.387 ≥0.25 Treatment 35
Richness
28 Alpha.Score Treatment Exposed Control −0.228 0.223 −1.021 ≥0.25 Treatment 28
32 Alpha.Score Treatment Exposed Control 0.601 0.132 4.546 <0.001 Treatment 32
35 Alpha.Score Treatment Exposed Control 0.463 0.092 5.021 <0.001 Treatment 35

5C)

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Capscale
ADONIS2
adonis2(Beta Distance ~ Temperature*Treatment); Pre-Exposed fish
term df SumOfSqs R2 statistic p.value sig
bray
Temperature 2.000 0.871 0.188 7.178 0.001 ***
Treatment 1.000 0.244 0.053 4.025 0.010 **
Temperature:Treatment 2.000 0.245 0.053 2.019 0.053 ns
Residual 54.000 3.275 0.707 NA NA NA
Total 59.000 4.635 1.000 NA NA NA
canberra
Temperature 2.000 1.652 0.132 4.615 0.001 ***
Treatment 1.000 0.533 0.043 2.979 0.001 ***
Temperature:Treatment 2.000 0.667 0.053 1.864 0.001 ***
Residual 54.000 9.665 0.772 NA NA NA
Total 59.000 12.517 1.000 NA NA NA
gunifrac
Temperature 2.000 0.647 0.153 5.604 0.001 ***
Treatment 1.000 0.288 0.068 4.987 0.003 **
Temperature:Treatment 2.000 0.177 0.042 1.537 0.109 ns
Residual 54.000 3.117 0.737 NA NA NA
Total 59.000 4.229 1.000 NA NA NA
Dispersion (ANOVA)
ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature); Pre-Exposed fish
term df sumsq meansq statistic p.value sig
bray
Temp.Treat 5.000 0.103 0.021 2.738 0.028 *
Residual 54.000 0.405 0.007 NA NA NA
canberra
Temp.Treat 5.000 0.014 0.003 0.453 ≥0.25 ns
Residual 54.000 0.329 0.006 NA NA NA
gunifrac
Temp.Treat 5.000 0.054 0.011 1.564 0.186 ns
Residual 54.000 0.369 0.007 NA NA NA
Dispersion (Tukey)
Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature*Treatment); Pre-Exposed fish
.y. term group1 group2 estimate conf.low conf.high adj.p.value sig
bray
Distance Temp.Treat 28°C__Exposed 28°C__Control 0.027 −0.088 0.141 ≥0.25 ns
Distance Temp.Treat 32°C__Control 28°C__Control 0.095 −0.019 0.210 0.153 ns
Distance Temp.Treat 32°C__Exposed 28°C__Control 0.110 −0.004 0.225 0.065 ns
Distance Temp.Treat 35°C__Control 28°C__Control 0.015 −0.099 0.130 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 28°C__Control 0.069 −0.045 0.183 ≥0.25 ns
Distance Temp.Treat 32°C__Control 28°C__Exposed 0.069 −0.046 0.183 ≥0.25 ns
Distance Temp.Treat 32°C__Exposed 28°C__Exposed 0.084 −0.031 0.198 ≥0.25 ns
Distance Temp.Treat 35°C__Control 28°C__Exposed −0.011 −0.126 0.103 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 28°C__Exposed 0.042 −0.072 0.157 ≥0.25 ns
Distance Temp.Treat 32°C__Exposed 32°C__Control 0.015 −0.100 0.129 ≥0.25 ns
Distance Temp.Treat 35°C__Control 32°C__Control −0.080 −0.195 0.034 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 32°C__Control −0.026 −0.141 0.088 ≥0.25 ns
Distance Temp.Treat 35°C__Control 32°C__Exposed −0.095 −0.209 0.019 0.156 ns
Distance Temp.Treat 35°C__Exposed 32°C__Exposed −0.041 −0.156 0.073 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 35°C__Control 0.054 −0.061 0.168 ≥0.25 ns
canberra
Distance Temp.Treat 28°C__Exposed 28°C__Control 0.011 −0.092 0.114 ≥0.25 ns
Distance Temp.Treat 32°C__Control 28°C__Control 0.035 −0.068 0.138 ≥0.25 ns
Distance Temp.Treat 32°C__Exposed 28°C__Control 0.044 −0.059 0.147 ≥0.25 ns
Distance Temp.Treat 35°C__Control 28°C__Control 0.010 −0.094 0.113 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 28°C__Control 0.016 −0.087 0.119 ≥0.25 ns
Distance Temp.Treat 32°C__Control 28°C__Exposed 0.024 −0.079 0.127 ≥0.25 ns
Distance Temp.Treat 32°C__Exposed 28°C__Exposed 0.033 −0.070 0.136 ≥0.25 ns
Distance Temp.Treat 35°C__Control 28°C__Exposed −0.001 −0.104 0.102 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 28°C__Exposed 0.005 −0.098 0.108 ≥0.25 ns
Distance Temp.Treat 32°C__Exposed 32°C__Control 0.009 −0.094 0.112 ≥0.25 ns
Distance Temp.Treat 35°C__Control 32°C__Control −0.025 −0.128 0.078 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 32°C__Control −0.019 −0.122 0.084 ≥0.25 ns
Distance Temp.Treat 35°C__Control 32°C__Exposed −0.034 −0.137 0.069 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 32°C__Exposed −0.028 −0.131 0.075 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 35°C__Control 0.006 −0.097 0.109 ≥0.25 ns
gunifrac
Distance Temp.Treat 28°C__Exposed 28°C__Control 0.003 −0.106 0.112 ≥0.25 ns
Distance Temp.Treat 32°C__Control 28°C__Control 0.051 −0.058 0.160 ≥0.25 ns
Distance Temp.Treat 32°C__Exposed 28°C__Control 0.078 −0.032 0.187 ≥0.25 ns
Distance Temp.Treat 35°C__Control 28°C__Control 0.000 −0.109 0.110 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 28°C__Control 0.042 −0.067 0.151 ≥0.25 ns
Distance Temp.Treat 32°C__Control 28°C__Exposed 0.048 −0.061 0.158 ≥0.25 ns
Distance Temp.Treat 32°C__Exposed 28°C__Exposed 0.075 −0.035 0.184 ≥0.25 ns
Distance Temp.Treat 35°C__Control 28°C__Exposed −0.002 −0.112 0.107 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 28°C__Exposed 0.039 −0.070 0.148 ≥0.25 ns
Distance Temp.Treat 32°C__Exposed 32°C__Control 0.026 −0.083 0.136 ≥0.25 ns
Distance Temp.Treat 35°C__Control 32°C__Control −0.051 −0.160 0.059 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 32°C__Control −0.009 −0.118 0.100 ≥0.25 ns
Distance Temp.Treat 35°C__Control 32°C__Exposed −0.077 −0.186 0.032 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 32°C__Exposed −0.036 −0.145 0.074 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 35°C__Control 0.041 −0.068 0.151 ≥0.25 ns

5D)

Tables
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Capscale
ADONIS2
adonis2(Beta Distance ~ Temperature*Treatment); Post-Exposed fish
term df SumOfSqs R2 statistic p.value sig
bray
Temperature 2.000 4.123 0.108 17.035 0.001 ***
Treatment 1.000 7.411 0.194 61.235 0.001 ***
Temperature:Treatment 2.000 3.168 0.083 13.088 0.001 ***
Residual 194.000 23.478 0.615 NA NA NA
Total 199.000 38.179 1.000 NA NA NA
canberra
Temperature 2.000 4.561 0.074 9.502 0.001 ***
Treatment 1.000 7.402 0.120 30.840 0.001 ***
Temperature:Treatment 2.000 3.014 0.049 6.279 0.001 ***
Residual 194.000 46.561 0.757 NA NA NA
Total 199.000 61.538 1.000 NA NA NA
gunifrac
Temperature 2.000 2.273 0.082 11.945 0.001 ***
Treatment 1.000 4.718 0.170 49.577 0.001 ***
Temperature:Treatment 2.000 2.273 0.082 11.944 0.001 ***
Residual 194.000 18.460 0.666 NA NA NA
Total 199.000 27.724 1.000 NA NA NA
Dispersion (ANOVA)
ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature); Post-Exposed fish
term df sumsq meansq statistic p.value sig
bray
Temp.Treat 5.000 1.344 0.269 17.882 <0.001 ****
Residual 194.000 2.917 0.015 NA NA NA
canberra
Temp.Treat 5.000 1.094 0.219 31.758 <0.001 ****
Residual 194.000 1.337 0.007 NA NA NA
gunifrac
Temp.Treat 5.000 1.225 0.245 29.264 <0.001 ****
Residual 194.000 1.624 0.008 NA NA NA
Dispersion (Tukey)
Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature*Treatment); Post-Exposed fish
.y. term group1 group2 estimate conf.low conf.high adj.p.value sig
bray
Distance Temp.Treat 28°C__Exposed 28°C__Control 0.211 0.119 0.302 <0.001 ****
Distance Temp.Treat 32°C__Control 28°C__Control −0.015 −0.096 0.066 ≥0.25 ns
Distance Temp.Treat 32°C__Exposed 28°C__Control 0.168 0.081 0.255 <0.001 ****
Distance Temp.Treat 35°C__Control 28°C__Control 0.013 −0.070 0.096 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 28°C__Control 0.060 −0.024 0.144 ≥0.25 ns
Distance Temp.Treat 32°C__Control 28°C__Exposed −0.225 −0.316 −0.135 <0.001 ****
Distance Temp.Treat 32°C__Exposed 28°C__Exposed −0.043 −0.138 0.053 ≥0.25 ns
Distance Temp.Treat 35°C__Control 28°C__Exposed −0.197 −0.290 −0.105 <0.001 ****
Distance Temp.Treat 35°C__Exposed 28°C__Exposed −0.150 −0.243 −0.057 <0.001 ****
Distance Temp.Treat 32°C__Exposed 32°C__Control 0.183 0.097 0.268 <0.001 ****
Distance Temp.Treat 35°C__Control 32°C__Control 0.028 −0.054 0.110 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 32°C__Control 0.075 −0.008 0.158 0.100 ns
Distance Temp.Treat 35°C__Control 32°C__Exposed −0.155 −0.243 −0.067 <0.001 ****
Distance Temp.Treat 35°C__Exposed 32°C__Exposed −0.108 −0.196 −0.019 0.007 **
Distance Temp.Treat 35°C__Exposed 35°C__Control 0.047 −0.038 0.132 ≥0.25 ns
canberra
Distance Temp.Treat 28°C__Exposed 28°C__Control 0.217 0.155 0.279 <0.001 ****
Distance Temp.Treat 32°C__Control 28°C__Control −0.011 −0.066 0.044 ≥0.25 ns
Distance Temp.Treat 32°C__Exposed 28°C__Control 0.107 0.048 0.166 <0.001 ****
Distance Temp.Treat 35°C__Control 28°C__Control 0.013 −0.044 0.069 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 28°C__Control 0.073 0.016 0.130 0.004 **
Distance Temp.Treat 32°C__Control 28°C__Exposed −0.228 −0.289 −0.167 <0.001 ****
Distance Temp.Treat 32°C__Exposed 28°C__Exposed −0.110 −0.175 −0.045 <0.001 ****
Distance Temp.Treat 35°C__Control 28°C__Exposed −0.204 −0.267 −0.142 <0.001 ****
Distance Temp.Treat 35°C__Exposed 28°C__Exposed −0.144 −0.207 −0.081 <0.001 ****
Distance Temp.Treat 32°C__Exposed 32°C__Control 0.118 0.060 0.176 <0.001 ****
Distance Temp.Treat 35°C__Control 32°C__Control 0.024 −0.032 0.080 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 32°C__Control 0.084 0.028 0.140 <0.001 ***
Distance Temp.Treat 35°C__Control 32°C__Exposed −0.094 −0.154 −0.035 <0.001 ***
Distance Temp.Treat 35°C__Exposed 32°C__Exposed −0.034 −0.094 0.026 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 35°C__Control 0.060 0.003 0.118 0.035 *
gunifrac
Distance Temp.Treat 28°C__Exposed 28°C__Control 0.197 0.129 0.266 <0.001 ****
Distance Temp.Treat 32°C__Control 28°C__Control −0.023 −0.083 0.037 ≥0.25 ns
Distance Temp.Treat 32°C__Exposed 28°C__Control 0.141 0.077 0.206 <0.001 ****
Distance Temp.Treat 35°C__Control 28°C__Control −0.011 −0.073 0.051 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 28°C__Control 0.054 −0.009 0.117 0.134 ns
Distance Temp.Treat 32°C__Control 28°C__Exposed −0.220 −0.288 −0.153 <0.001 ****
Distance Temp.Treat 32°C__Exposed 28°C__Exposed −0.056 −0.128 0.015 0.211 ns
Distance Temp.Treat 35°C__Control 28°C__Exposed −0.209 −0.278 −0.140 <0.001 ****
Distance Temp.Treat 35°C__Exposed 28°C__Exposed −0.143 −0.213 −0.074 <0.001 ****
Distance Temp.Treat 32°C__Exposed 32°C__Control 0.164 0.100 0.228 <0.001 ****
Distance Temp.Treat 35°C__Control 32°C__Control 0.012 −0.050 0.073 ≥0.25 ns
Distance Temp.Treat 35°C__Exposed 32°C__Control 0.077 0.015 0.139 0.006 **
Distance Temp.Treat 35°C__Control 32°C__Exposed −0.152 −0.218 −0.087 <0.001 ****
Distance Temp.Treat 35°C__Exposed 32°C__Exposed −0.087 −0.153 −0.021 0.003 **
Distance Temp.Treat 35°C__Exposed 35°C__Control 0.065 0.002 0.129 0.039 *

6) Water temperature moderates the impact of infection burden on fish gut microbiomes

Fig. 6 Exposure to Pseudocapillaria tomentosa differentially impacts gut microbiome depending on water temperature. (A) Simpson’s Index for diversity of parasite exposed fish. Gut microbial alpha-diversity does not significantly differ between fish reared at the same water temperature with presence or absence of infection. (B) Capscale ordination based on the Canberra dissimilarity of gut microbiome composition of parasite exposed fish. The analysis shows that gut microbiome composition significantly differs between positively infected fish reared at different water temperatures. (C) Total worm count is positively correlated with lowest or highest alpha diversity scores in fish with detectable parasites. Points are colored by temperature and filled by cluster (High = purple, Low = orange, Other = white), and samples with a worm count equal to 0 are transparent. (D) Capscale ordination based on the Bray-Curtis dissimilarity of gut microbiome composition of water temperature and worm counts. Points are colored by temperature and filled by cluster (High = purple, Low = orange, Other = white), and samples with a worm count equal to 0 are transparent. The analysis shows that gut microbiome composition significantly differs between water temperature and total worm count, and samples labeled as “High” or “Low” form distinct clusters. Ribbons and ellipses indicate 95% confidence interval. Only statistically significant relationships are shown. A “*” indicates statistical significance below the “0.05” level. Black arrows indicate statistically significant covariates and direction of greatest change in the indicated covariates.

6A)

Tables
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GLM
GLM Results
glm(Alpha.Score ~ Temperature*Pathology.Results); Exposed fish
term estimate std.error statistic p.value p.adj.sig
Shannon
(Intercept) −1.090 0.429 −2.544 0.013 *
Temperature32 0.889 0.508 1.749 0.084 ns
Temperature35 0.693 0.468 1.482 0.142 ns
Pathology.Resultspositive 0.223 0.512 0.436 ≥0.25 ns
Temperature32:Pathology.Resultspositive 0.125 0.641 0.195 ≥0.25 ns
Temperature35:Pathology.Resultspositive −1.122 1.392 −0.806 ≥0.25 ns
Simpson
(Intercept) −1.265 0.458 −2.760 0.007 **
Temperature32 1.105 0.536 2.061 0.042 *
Temperature35 1.055 0.495 2.130 0.036 *
Pathology.Resultspositive 0.375 0.541 0.694 ≥0.25 ns
Temperature32:Pathology.Resultspositive 0.054 0.669 0.081 ≥0.25 ns
Temperature35:Pathology.Resultspositive −1.154 1.337 −0.863 ≥0.25 ns
Richness
(Intercept) −0.421 0.329 −1.279 0.204 ns
Temperature32 0.483 0.405 1.194 0.236 ns
Temperature35 −0.002 0.367 −0.005 ≥0.25 ns
Pathology.Resultspositive −0.390 0.407 −0.957 ≥0.25 ns
Temperature32:Pathology.Resultspositive 0.310 0.526 0.589 ≥0.25 ns
Temperature35:Pathology.Resultspositive −0.221 1.125 −0.197 ≥0.25 ns
ANOVA
ANOVA of GLM
ANOVA(GLM(Alpha.Score ~ Temperature*Time), type = 2); Exposed fish
term statistic df p.value sig
Shannon
Temperature 9.895 2.000 0.007 **
Pathology.Results 0.569 1.000 ≥0.25 ns
Temperature:Pathology.Results 0.997 2.000 ≥0.25 ns
Simpson
Temperature 13.663 2.000 0.001 **
Pathology.Results 1.134 1.000 ≥0.25 ns
Temperature:Pathology.Results 0.999 2.000 ≥0.25 ns
Richness
Temperature 8.625 2.000 0.013 *
Pathology.Results 0.840 1.000 ≥0.25 ns
Temperature:Pathology.Results 0.495 2.000 ≥0.25 ns
Tukey
Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Alpha.Score ~ Temperature*Time); Exposed fish
Temperature .y. term group1 group2 estimate std.error statistic adj.p.value Variable Group
Shannon
28 Alpha.Score Pathology.Results positive negative 0.223 0.732 0.305 ≥0.25 Pathology.Results 28
32 Alpha.Score Pathology.Results positive negative 0.348 0.383 0.908 ≥0.25 Pathology.Results 32
35 Alpha.Score Pathology.Results positive negative −0.899 0.659 −1.365 0.172 Pathology.Results 35
Simpson
28 Alpha.Score Pathology.Results positive negative 0.375 0.754 0.497 ≥0.25 Pathology.Results 28
32 Alpha.Score Pathology.Results positive negative 0.429 0.394 1.089 ≥0.25 Pathology.Results 32
35 Alpha.Score Pathology.Results positive negative −0.779 0.692 −1.125 ≥0.25 Pathology.Results 35
Richness
28 Alpha.Score Pathology.Results positive negative −0.390 0.608 −0.641 ≥0.25 Pathology.Results 28
32 Alpha.Score Pathology.Results positive negative −0.080 0.301 −0.267 ≥0.25 Pathology.Results 32
35 Alpha.Score Pathology.Results positive negative −0.611 0.552 −1.107 ≥0.25 Pathology.Results 35

6B)

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Capscale
ADONIS2
adonis2(Beta Distance ~ Temperature*Pathology.Results); Exposed fish
term df SumOfSqs R2 statistic p.value sig
bray
Temperature 2.000 5.218 0.251 14.569 0.001 ***
Pathology.Results 1.000 0.263 0.013 1.471 0.185 ns
Temperature:Pathology.Results 2.000 0.404 0.019 1.127 ≥0.25 ns
Residual 83.000 14.863 0.716 NA NA NA
Total 88.000 20.748 1.000 NA NA NA
canberra
Temperature 2.000 3.573 0.117 5.751 0.001 ***
Pathology.Results 1.000 0.488 0.016 1.570 0.031 *
Temperature:Pathology.Results 2.000 0.679 0.022 1.093 ≥0.25 ns
Residual 83.000 25.781 0.845 NA NA NA
Total 88.000 30.520 1.000 NA NA NA
gunifrac
Temperature 2.000 3.363 0.212 11.651 0.001 ***
Pathology.Results 1.000 0.234 0.015 1.622 0.149 ns
Temperature:Pathology.Results 2.000 0.264 0.017 0.915 ≥0.25 ns
Residual 83.000 11.980 0.756 NA NA NA
Total 88.000 15.841 1.000 NA NA NA
Dispersion (ANOVA)
ANOVA: Homogeneity of Dispersion
ANOVA(Beta Disperson ~ Temperature*Pathology.Results); Exposed fish
term df sumsq meansq statistic p.value sig
bray
Temp.Path 5.000 0.520 0.104 4.652 <0.001 ***
Residual 83.000 1.855 0.022 NA NA NA
canberra
Temp.Path 5.000 0.625 0.125 13.830 <0.001 ****
Residual 83.000 0.750 0.009 NA NA NA
gunifrac
Temp.Path 5.000 0.436 0.087 6.954 <0.001 ****
Residual 83.000 1.040 0.013 NA NA NA
Dispersion (Tukey)
Tukey: Homogeneity of Dispersion
Tukey(Beta Disperson ~ Temperature*Pathology.Results); Exposed fish
.y. term group1 group2 estimate conf.low conf.high adj.p.value sig
bray
Distance Temp.Path 28°C__pos 28°C__neg 0.022 −0.165 0.209 ≥0.25 ns
Distance Temp.Path 32°C__neg 28°C__neg −0.086 −0.277 0.105 ≥0.25 ns
Distance Temp.Path 32°C__pos 28°C__neg 0.015 −0.176 0.206 ≥0.25 ns
Distance Temp.Path 35°C__neg 28°C__neg −0.126 −0.298 0.046 ≥0.25 ns
Distance Temp.Path 35°C__pos 28°C__neg −0.434 −0.896 0.029 0.079 ns
Distance Temp.Path 32°C__neg 28°C__pos −0.108 −0.263 0.046 ≥0.25 ns
Distance Temp.Path 32°C__pos 28°C__pos −0.007 −0.161 0.148 ≥0.25 ns
Distance Temp.Path 35°C__neg 28°C__pos −0.148 −0.278 −0.017 0.017 *
Distance Temp.Path 35°C__pos 28°C__pos −0.456 −0.905 −0.007 0.044 *
Distance Temp.Path 32°C__pos 32°C__neg 0.101 −0.058 0.261 ≥0.25 ns
Distance Temp.Path 35°C__neg 32°C__neg −0.039 −0.175 0.096 ≥0.25 ns
Distance Temp.Path 35°C__pos 32°C__neg −0.348 −0.798 0.103 0.226 ns
Distance Temp.Path 35°C__neg 32°C__pos −0.141 −0.277 −0.005 0.038 *
Distance Temp.Path 35°C__pos 32°C__pos −0.449 −0.899 0.001 0.051 ns
Distance Temp.Path 35°C__pos 35°C__neg −0.308 −0.751 0.135 ≥0.25 ns
canberra
Distance Temp.Path 28°C__pos 28°C__neg 0.045 −0.074 0.164 ≥0.25 ns
Distance Temp.Path 32°C__neg 28°C__neg −0.123 −0.244 −0.002 0.045 *
Distance Temp.Path 32°C__pos 28°C__neg −0.042 −0.164 0.079 ≥0.25 ns
Distance Temp.Path 35°C__neg 28°C__neg −0.105 −0.214 0.005 0.068 ns
Distance Temp.Path 35°C__pos 28°C__neg −0.588 −0.883 −0.294 <0.001 ****
Distance Temp.Path 32°C__neg 28°C__pos −0.168 −0.267 −0.070 <0.001 ****
Distance Temp.Path 32°C__pos 28°C__pos −0.088 −0.186 0.010 0.107 ns
Distance Temp.Path 35°C__neg 28°C__pos −0.150 −0.233 −0.067 <0.001 ****
Distance Temp.Path 35°C__pos 28°C__pos −0.634 −0.919 −0.348 <0.001 ****
Distance Temp.Path 32°C__pos 32°C__neg 0.080 −0.021 0.182 0.199 ns
Distance Temp.Path 35°C__neg 32°C__neg 0.018 −0.068 0.105 ≥0.25 ns
Distance Temp.Path 35°C__pos 32°C__neg −0.465 −0.752 −0.179 <0.001 ***
Distance Temp.Path 35°C__neg 32°C__pos −0.062 −0.149 0.024 ≥0.25 ns
Distance Temp.Path 35°C__pos 32°C__pos −0.546 −0.832 −0.259 <0.001 ****
Distance Temp.Path 35°C__pos 35°C__neg −0.484 −0.765 −0.202 <0.001 ****
gunifrac
Distance Temp.Path 28°C__pos 28°C__neg 0.011 −0.129 0.151 ≥0.25 ns
Distance Temp.Path 32°C__neg 28°C__neg −0.091 −0.234 0.052 ≥0.25 ns
Distance Temp.Path 32°C__pos 28°C__neg −0.014 −0.157 0.129 ≥0.25 ns
Distance Temp.Path 35°C__neg 28°C__neg −0.129 −0.258 0.000 0.049 *
Distance Temp.Path 35°C__pos 28°C__neg −0.411 −0.758 −0.065 0.011 *
Distance Temp.Path 32°C__neg 28°C__pos −0.102 −0.218 0.014 0.115 ns
Distance Temp.Path 32°C__pos 28°C__pos −0.025 −0.141 0.091 ≥0.25 ns
Distance Temp.Path 35°C__neg 28°C__pos −0.140 −0.238 −0.043 <0.001 ***
Distance Temp.Path 35°C__pos 28°C__pos −0.423 −0.759 −0.087 0.006 **
Distance Temp.Path 32°C__pos 32°C__neg 0.077 −0.042 0.196 ≥0.25 ns
Distance Temp.Path 35°C__neg 32°C__neg −0.038 −0.140 0.064 ≥0.25 ns
Distance Temp.Path 35°C__pos 32°C__neg −0.321 −0.658 0.017 0.072 ns
Distance Temp.Path 35°C__neg 32°C__pos −0.115 −0.217 −0.013 0.017 *
Distance Temp.Path 35°C__pos 32°C__pos −0.398 −0.735 −0.060 0.011 *
Distance Temp.Path 35°C__pos 35°C__neg −0.283 −0.614 0.049 0.140 ns

6C)

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GLM
GLM Results
glm(Total.Worm.Count ~ Alpha.Score*Cluster); Exposed fish
term estimate std.error statistic p.value p.adj.sig
Shannon
(Intercept) 0.000 0.778 0.000 ≥0.25 ns
Alpha.Score 0.000 1.808 0.000 ≥0.25 ns
ClusterLow 6.524 1.067 6.115 <0.001 ****
ClusterHigh −2.610 3.512 −0.743 ≥0.25 ns
Alpha.Score:ClusterLow −10.090 4.999 −2.018 0.047 *
Alpha.Score:ClusterHigh 13.706 4.766 2.876 0.005 **
Simpson
(Intercept) 0.000 0.741 0.000 ≥0.25 ns
Alpha.Score 0.000 1.605 0.000 ≥0.25 ns
ClusterLow 6.842 1.041 6.573 <0.001 ****
ClusterHigh −3.247 3.900 −0.833 ≥0.25 ns
Alpha.Score:ClusterLow −12.165 4.647 −2.618 0.011 *
Alpha.Score:ClusterHigh 14.198 5.090 2.789 0.007 **
Richness
(Intercept) 0.000 0.914 0.000 ≥0.25 ns
Alpha.Score 0.000 1.983 0.000 ≥0.25 ns
ClusterLow 6.105 1.294 4.717 <0.001 ****
ClusterHigh 7.362 2.644 2.784 0.007 **
Alpha.Score:ClusterLow −3.491 4.463 −0.782 ≥0.25 ns
Alpha.Score:ClusterHigh 0.772 4.163 0.185 ≥0.25 ns
ANOVA
ANOVA of GLM
ANOVA(GLM(Total.Worm.Count ~ Alpha.Score*Cluster), type = 2); Exposed fish
term statistic df p.value sig
Shannon
Alpha.Score 0.143 1.000 ≥0.25 ns
Cluster 162.212 2.000 <0.001 ****
Alpha.Score:Cluster 14.204 2.000 <0.001 ***
Simpson
Alpha.Score 0.002 1.000 ≥0.25 ns
Cluster 168.513 2.000 <0.001 ****
Alpha.Score:Cluster 16.419 2.000 <0.001 ***
Richness
Alpha.Score 0.066 1.000 ≥0.25 ns
Cluster 145.539 2.000 <0.001 ****
Alpha.Score:Cluster 0.741 2.000 ≥0.25 ns
Tukey
Pairwise Tukey's HSD, p.adj: Dunnett
Tukey(Total.Worm.Count ~ Alpha.Score*Cluster); Exposed fish
Temperature .y. term group1 group2 estimate std.error statistic adj.p.value Variable Group
Shannon
28 Alpha.Score Cluster Low Other −1.594 0.642 −2.483 0.034 Cluster 28
28 Alpha.Score Cluster High Other 2.844 0.672 4.233 <0.001 Cluster 28
28 Alpha.Score Cluster High Low 4.438 0.773 5.741 <0.001 Cluster 28
32 Alpha.Score Cluster Low Other −1.391 0.495 −2.808 0.013 Cluster 32
32 Alpha.Score Cluster High Other 1.147 0.333 3.443 0.002 Cluster 32
32 Alpha.Score Cluster High Low 2.538 0.528 4.811 <0.001 Cluster 32
35 Alpha.Score Cluster Low Other −0.899 0.659 −1.365 0.172 Cluster 35
Simpson
28 Alpha.Score Cluster Low Other −1.487 0.656 −2.266 0.060 Cluster 28
28 Alpha.Score Cluster High Other 2.970 0.665 4.466 <0.001 Cluster 28
28 Alpha.Score Cluster High Low 4.457 0.767 5.810 <0.001 Cluster 28
32 Alpha.Score Cluster Low Other −1.233 0.487 −2.531 0.029 Cluster 32
32 Alpha.Score Cluster High Other 1.264 0.354 3.573 <0.001 Cluster 32
32 Alpha.Score Cluster High Low 2.497 0.528 4.729 <0.001 Cluster 32
35 Alpha.Score Cluster Low Other −0.779 0.692 −1.125 ≥0.25 Cluster 35
Richness
28 Alpha.Score Cluster Low Other −1.430 0.589 −2.429 0.040 Cluster 28
28 Alpha.Score Cluster High Other 1.367 0.653 2.095 0.090 Cluster 28
28 Alpha.Score Cluster High Low 2.797 0.692 4.043 <0.001 Cluster 28
32 Alpha.Score Cluster Low Other −1.183 0.387 −3.052 0.006 Cluster 32
32 Alpha.Score Cluster High Other 0.429 0.278 1.543 ≥0.25 Cluster 32
32 Alpha.Score Cluster High Low 1.611 0.410 3.934 <0.001 Cluster 32
35 Alpha.Score Cluster Low Other −0.611 0.552 −1.107 ≥0.25 Cluster 35

6D)

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Capscale
Dispersion (ANOVA)
Dispersion (Tukey)

7) Gut microbial abundance is significantly associated with environmental conditions and stressors

7A)

Fig. 7 A heatmap of model coefficient values of the top 50 statistically significant abundant gut microbial taxa identified by MaAsLin2. The color of each cell represents the coefficient value and direction (red is positive, blue is negative). A “+” or “-” indicates a statistically significant association was observed between taxon abundance and a covariate. Gray colored cells indicate a significant effect was not observed.

Taxon Temp: 32°C Temp: 35°C Time (DPE) Cluster: Low Parasite exposed Infection present Infection burden
Actinobacteriota
IMCC26207 +
- - -

Nocardiaceae Genus + + +



PeM15 Genus +
- - -

Bacteroidota
Barnesiellaceae Genus

-
-

Cloacibacterium
+ -



Flavobacterium - - - - +

Fluviicola
- -
- -
Microscillaceae Genus + + + -


Terrimonas


-
+ +
env OPS 17 Genus
-
- -

Chloroflexi
JG30 KF CM45 Genus

+ - + +
Cyanobacteria
Candidatus Obscuribacter - - + - + +
Obscuribacteraceae Genus +
+ -
+ -
Vampirovibrionaceae Genus

+ - + +
Desulfobacterota
Desulfobacterota Genus


- + +
Firmicutes
Clostridium sensu stricto 3

+ - + +
Gemmatimonadota
Gemmatimonadaceae Genus
-
- + +
Myxococcota
Sandaracinaceae Genus + + + - + +
NB1-j
NB1 j Genus
+ + - + +
Nitrospirota
Nitrospira
- + - + +
Patescibacteria
Saccharimonadales Genus + + + - +

Planctomycetota
CL500 3

+ - - +
Gemmata
+ + - + +
Pirellulaceae Genus + - + - -

Planctomycetales Genus

+ - + +
Planctomycetes Genus

+ - + +
Rhodopirellula


- + + -
SM1A02

+ - + +
Telmatocola

+ - + +
Proteobacteria
A0839 Genus +

- + +
Alphaproteobacteria Genus
+ + - + +
Bauldia +
+ - + +
Beijerinckiaceae Genus
- + -
+ -
Bosea - - - -


Candidatus Alysiosphaera +
+ - + +
Chitinibacter
+ +
+
-
Comamonadaceae Genus

- - + +
Gammaproteobacteria Genus - - + -
+
Luteimonas -

-
+ +
Methyloligellaceae Genus + + + - + +
Nordella
-
- + +
Plesiomonas + +

- - -
Pseudomonadales Genus - - - - - -
Rhizobacter


- + + +
Rhizobiales Incertae Sedis Genus


-
+
Rhodobacteraceae Genus


- + +
Rhodovastum
-
-


Steroidobacteraceae Genus

+ - + +
Xanthobacteraceae Genus - - + -
+
Verrucomicrobiota
Oikopleura

+ - + +